A technique for identifying specific DNA sequences that are bound, in vivo, to proteins of interest. It involves formaldehyde fixation of CHROMATIN to crosslink the DNA-BINDING PROTEINS to the DNA. After shearing the DNA into small fragments, specific DNA-protein complexes are isolated by immunoprecipitation with protein-specific ANTIBODIES. Then, the DNA isolated from the complex can be identified by PCR amplification and sequencing.
The aggregation of soluble ANTIGENS with ANTIBODIES, alone or with antibody binding factors such as ANTI-ANTIBODIES or STAPHYLOCOCCAL PROTEIN A, into complexes large enough to fall out of solution.
Serologic tests in which a positive reaction manifested by visible CHEMICAL PRECIPITATION occurs when a soluble ANTIGEN reacts with its precipitins, i.e., ANTIBODIES that can form a precipitate.
DNA sequences which are recognized (directly or indirectly) and bound by a DNA-dependent RNA polymerase during the initiation of transcription. Highly conserved sequences within the promoter include the Pribnow box in bacteria and the TATA BOX in eukaryotes.
The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.
Established cell cultures that have the potential to propagate indefinitely.
Descriptions of specific amino acid, carbohydrate, or nucleotide sequences which have appeared in the published literature and/or are deposited in and maintained by databanks such as GENBANK, European Molecular Biology Laboratory (EMBL), National Biomedical Research Foundation (NBRF), or other sequence repositories.
The biosynthesis of RNA carried out on a template of DNA. The biosynthesis of DNA from an RNA template is called REVERSE TRANSCRIPTION.
The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.
The material of CHROMOSOMES. It is a complex of DNA; HISTONES; and nonhistone proteins (CHROMOSOMAL PROTEINS, NON-HISTONE) found within the nucleus of a cell.
Endogenous substances, usually proteins, which are effective in the initiation, stimulation, or termination of the genetic transcription process.
RNA sequences that serve as templates for protein synthesis. Bacterial mRNAs are generally primary transcripts in that they do not require post-transcriptional processing. Eukaryotic mRNA is synthesized in the nucleus and must be exported to the cytoplasm for translation. Most eukaryotic mRNAs have a sequence of polyadenylic acid at the 3' end, referred to as the poly(A) tail. The function of this tail is not known for certain, but it may play a role in the export of mature mRNA from the nucleus as well as in helping stabilize some mRNA molecules by retarding their degradation in the cytoplasm.
The parts of a macromolecule that directly participate in its specific combination with another molecule.
Small chromosomal proteins (approx 12-20 kD) possessing an open, unfolded structure and attached to the DNA in cell nuclei by ionic linkages. Classification into the various types (designated histone I, histone II, etc.) is based on the relative amounts of arginine and lysine in each.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control (induction or repression) of gene action at the level of transcription or translation.
Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others.
Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
The sum of the weight of all the atoms in a molecule.
A cell line derived from cultured tumor cells.
Proteins which bind to DNA. The family includes proteins which bind to both double- and single-stranded DNA and also includes specific DNA binding proteins in serum which can be used as markers for malignant diseases.
Nucleotide sequences, usually upstream, which are recognized by specific regulatory transcription factors, thereby causing gene response to various regulatory agents. These elements may be found in both promoter and enhancer regions.
The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION.
Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody.
The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.
Formation of an acetyl derivative. (Stedman, 25th ed)
The uptake of naked or purified DNA by CELLS, usually meaning the process as it occurs in eukaryotic cells. It is analogous to bacterial transformation (TRANSFORMATION, BACTERIAL) and both are routinely employed in GENE TRANSFER TECHNIQUES.
Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.
Processes that stimulate the GENETIC TRANSCRIPTION of a gene or set of genes.
An electrophoretic technique for assaying the binding of one compound to another. Typically one compound is labeled to follow its mobility during electrophoresis. If the labeled compound is bound by the other compound, then the mobility of the labeled compound through the electrophoretic medium will be retarded.
Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus.
Antibodies produced by a single clone of cells.
Proteins which maintain the transcriptional quiescence of specific GENES or OPERONS. Classical repressor proteins are DNA-binding proteins that are normally bound to the OPERATOR REGION of an operon, or the ENHANCER SEQUENCES of a gene until a signal occurs that causes their release.
The intracellular transfer of information (biological activation/inhibition) through a signal pathway. In each signal transduction system, an activation/inhibition signal from a biologically active molecule (hormone, neurotransmitter) is mediated via the coupling of a receptor/enzyme to a second messenger system or to an ion channel. Signal transduction plays an important role in activating cellular functions, cell differentiation, and cell proliferation. Examples of signal transduction systems are the GAMMA-AMINOBUTYRIC ACID-postsynaptic receptor-calcium ion channel system, the receptor-mediated T-cell activation pathway, and the receptor-mediated activation of phospholipases. Those coupled to membrane depolarization or intracellular release of calcium include the receptor-mediated activation of cytotoxic functions in granulocytes and the synaptic potentiation of protein kinase activation. Some signal transduction pathways may be part of larger signal transduction pathways; for example, protein kinase activation is part of the platelet activation signal pathway.
Promoter-specific RNA polymerase II transcription factor that binds to the GC box, one of the upstream promoter elements, in mammalian cells. The binding of Sp1 is necessary for the initiation of transcription in the promoters of a variety of cellular and viral GENES.
The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for VIRUS CULTIVATION and antitumor drug screening assays.
Immunologic method used for detecting or quantifying immunoreactive substances. The substance is identified by first immobilizing it by blotting onto a membrane and then tagging it with labeled antibodies.
Small double-stranded, non-protein coding RNAs (21-31 nucleotides) involved in GENE SILENCING functions, especially RNA INTERFERENCE (RNAi). Endogenously, siRNAs are generated from dsRNAs (RNA, DOUBLE-STRANDED) by the same ribonuclease, Dicer, that generates miRNAs (MICRORNAS). The perfect match of the siRNAs' antisense strand to their target RNAs mediates RNAi by siRNA-guided RNA cleavage. siRNAs fall into different classes including trans-acting siRNA (tasiRNA), repeat-associated RNA (rasiRNA), small-scan RNA (scnRNA), and Piwi protein-interacting RNA (piRNA) and have different specific gene silencing functions.
The formation of a solid in a solution as a result of a chemical reaction or the aggregation of soluble substances into complexes large enough to fall out of solution.
A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine).
Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.
A variation of the PCR technique in which cDNA is made from RNA via reverse transcription. The resultant cDNA is then amplified using standard PCR protocols.
Diffusible gene products that act on homologous or heterologous molecules of viral or cellular DNA to regulate the expression of proteins.
Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.
Any detectable and heritable change in the genetic material that causes a change in the GENOTYPE and which is transmitted to daughter cells and to succeeding generations.
Any of various enzymatically catalyzed post-translational modifications of PEPTIDES or PROTEINS in the cell of origin. These modifications include carboxylation; HYDROXYLATION; ACETYLATION; PHOSPHORYLATION; METHYLATION; GLYCOSYLATION; ubiquitination; oxidation; proteolysis; and crosslinking and result in changes in molecular weight and electrophoretic motility.
The biosynthesis of PEPTIDES and PROTEINS on RIBOSOMES, directed by MESSENGER RNA, via TRANSFER RNA that is charged with standard proteinogenic AMINO ACIDS.
Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)
Interruption or suppression of the expression of a gene at transcriptional or translational levels.
Deacetylases that remove N-acetyl groups from amino side chains of the amino acids of HISTONES. The enzyme family can be divided into at least three structurally-defined subclasses. Class I and class II deacetylases utilize a zinc-dependent mechanism. The sirtuin histone deacetylases belong to class III and are NAD-dependent enzymes.
Screening techniques first developed in yeast to identify genes encoding interacting proteins. Variations are used to evaluate interplay between proteins and other molecules. Two-hybrid techniques refer to analysis for protein-protein interactions, one-hybrid for DNA-protein interactions, three-hybrid interactions for RNA-protein interactions or ligand-based interactions. Reverse n-hybrid techniques refer to analysis for mutations or other small molecules that dissociate known interactions.
Recombinant proteins produced by the GENETIC TRANSLATION of fused genes formed by the combination of NUCLEIC ACID REGULATORY SEQUENCES of one or more genes with the protein coding sequences of one or more genes.
A gene silencing phenomenon whereby specific dsRNAs (RNA, DOUBLE-STRANDED) trigger the degradation of homologous mRNA (RNA, MESSENGER). The specific dsRNAs are processed into SMALL INTERFERING RNA (siRNA) which serves as a guide for cleavage of the homologous mRNA in the RNA-INDUCED SILENCING COMPLEX. DNA METHYLATION may also be triggered during this process.
Proteins prepared by recombinant DNA technology.
Transport proteins that carry specific substances in the blood or across cell membranes.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in neoplastic tissue.
The level of protein structure in which combinations of secondary protein structures (alpha helices, beta sheets, loop regions, and motifs) pack together to form folded shapes called domains. Disulfide bridges between cysteines in two different parts of the polypeptide chain along with other interactions between the chains play a role in the formation and stabilization of tertiary structure. Small proteins usually consist of only one domain but larger proteins may contain a number of domains connected by segments of polypeptide chain which lack regular secondary structure.
Immunoglobulin molecules having a specific amino acid sequence by virtue of which they interact only with the ANTIGEN (or a very similar shape) that induced their synthesis in cells of the lymphoid series (especially PLASMA CELLS).
Genes whose expression is easily detectable and therefore used to study promoter activity at many positions in a target genome. In recombinant DNA technology, these genes may be attached to a promoter region of interest.
Short sequences (generally about 10 base pairs) of DNA that are complementary to sequences of messenger RNA and allow reverse transcriptases to start copying the adjacent sequences of mRNA. Primers are used extensively in genetic and molecular biology techniques.
Cells grown in vitro from neoplastic tissue. If they can be established as a TUMOR CELL LINE, they can be propagated in cell culture indefinitely.
Sites on an antigen that interact with specific antibodies.
The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells.
Enzymes that oxidize certain LUMINESCENT AGENTS to emit light (PHYSICAL LUMINESCENCE). The luciferases from different organisms have evolved differently so have different structures and substrates.
The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site.
The artificial induction of GENE SILENCING by the use of RNA INTERFERENCE to reduce the expression of a specific gene. It includes the use of DOUBLE-STRANDED RNA, such as SMALL INTERFERING RNA and RNA containing HAIRPIN LOOP SEQUENCE, and ANTI-SENSE OLIGONUCLEOTIDES.
The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells.
A negative regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
Compounds and molecular complexes that consist of very large numbers of atoms and are generally over 500 kDa in size. In biological systems macromolecular substances usually can be visualized using ELECTRON MICROSCOPY and are distinguished from ORGANELLES by the lack of a membrane structure.
A positive regulatory effect on physiological processes at the molecular, cellular, or systemic level. At the molecular level, the major regulatory sites include membrane receptors, genes (GENE EXPRESSION REGULATION), mRNAs (RNA, MESSENGER), and proteins.
Hybridization of a nucleic acid sample to a very large set of OLIGONUCLEOTIDE PROBES, which have been attached individually in columns and rows to a solid support, to determine a BASE SEQUENCE, or to detect variations in a gene sequence, GENE EXPRESSION, or for GENE MAPPING.
Proteins found in any species of virus.
Addition of methyl groups to DNA. DNA methyltransferases (DNA methylases) perform this reaction using S-ADENOSYLMETHIONINE as the methyl group donor.
The rate dynamics in chemical or physical systems.
A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. It functions in the nucleoplasmic structure and transcribes DNA into RNA. It has different requirements for cations and salt than RNA polymerase I and is strongly inhibited by alpha-amanitin. EC 2.7.7.6.
The phenotypic manifestation of a gene or genes by the processes of GENETIC TRANSCRIPTION and GENETIC TRANSLATION.
Conjugated protein-carbohydrate compounds including mucins, mucoid, and amyloid glycoproteins.
Antigens on surfaces of cells, including infectious or foreign cells or viruses. They are usually protein-containing groups on cell membranes or walls and may be isolated.
A genetic process by which the adult organism is realized via mechanisms that lead to the restriction in the possible fates of cells, eventually leading to their differentiated state. Mechanisms involved cause heritable changes to cells without changes to DNA sequence such as DNA METHYLATION; HISTONE modification; DNA REPLICATION TIMING; NUCLEOSOME positioning; and heterochromatization which result in selective gene expression or repression.
Proteins encoded by homeobox genes (GENES, HOMEOBOX) that exhibit structural similarity to certain prokaryotic and eukaryotic DNA-binding proteins. Homeodomain proteins are involved in the control of gene expression during morphogenesis and development (GENE EXPRESSION REGULATION, DEVELOPMENTAL).
Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.
Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action in enzyme synthesis.
Progressive restriction of the developmental potential and increasing specialization of function that leads to the formation of specialized cells, tissues, and organs.
Proteins that bind to RNA molecules. Included here are RIBONUCLEOPROTEINS and other proteins whose function is to bind specifically to RNA.
CELL LINES derived from the CV-1 cell line by transformation with a replication origin defective mutant of SV40 VIRUS, which codes for wild type large T antigen (ANTIGENS, POLYOMAVIRUS TRANSFORMING). They are used for transfection and cloning. (The CV-1 cell line was derived from the kidney of an adult male African green monkey (CERCOPITHECUS AETHIOPS).)
Serum that contains antibodies. It is obtained from an animal that has been immunized either by ANTIGEN injection or infection with microorganisms containing the antigen.
Field of chemistry that pertains to immunological phenomena and the study of chemical reactions related to antigen stimulation of tissues. It includes physicochemical interactions between antigens and antibodies.
A species of the genus SACCHAROMYCES, family Saccharomycetaceae, order Saccharomycetales, known as "baker's" or "brewer's" yeast. The dried form is used as a dietary supplement.
A cell line generated from human embryonic kidney cells that were transformed with human adenovirus type 5.
Enzymes that catalyze acyl group transfer from ACETYL-CoA to HISTONES forming CoA and acetyl-histones.
A family of histone acetyltransferases that is structurally-related to CREB-BINDING PROTEIN and to E1A-ASSOCIATED P300 PROTEIN. They function as transcriptional coactivators by bridging between DNA-binding TRANSCRIPTION FACTORS and the basal transcription machinery. They also modify transcription factors and CHROMATIN through ACETYLATION.
Addition of methyl groups. In histo-chemistry methylation is used to esterify carboxyl groups and remove sulfate groups by treating tissue sections with hot methanol in the presence of hydrochloric acid. (From Stedman, 25th ed)
The species Oryctolagus cuniculus, in the family Leporidae, order LAGOMORPHA. Rabbits are born in burrows, furless, and with eyes and ears closed. In contrast with HARES, rabbits have 22 chromosome pairs.
A class of weak acids with the general formula R-CONHOH.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
Unstable isotopes of sulfur that decay or disintegrate spontaneously emitting radiation. S 29-31, 35, 37, and 38 are radioactive sulfur isotopes.
The process of moving proteins from one cellular compartment (including extracellular) to another by various sorting and transport mechanisms such as gated transport, protein translocation, and vesicular transport.
A heterotrimeric DNA-binding protein that binds to CCAAT motifs in the promoters of eukaryotic genes. It is composed of three subunits: A, B and C.
A subfamily in the family MURIDAE, comprising the hamsters. Four of the more common genera are Cricetus, CRICETULUS; MESOCRICETUS; and PHODOPUS.
Linear POLYPEPTIDES that are synthesized on RIBOSOMES and may be further modified, crosslinked, cleaved, or assembled into complex proteins with several subunits. The specific sequence of AMINO ACIDS determines the shape the polypeptide will take, during PROTEIN FOLDING, and the function of the protein.
Serological reactions in which an antiserum against one antigen reacts with a non-identical but closely related antigen.
A species of CERCOPITHECUS containing three subspecies: C. tantalus, C. pygerythrus, and C. sabeus. They are found in the forests and savannah of Africa. The African green monkey (C. pygerythrus) is the natural host of SIMIAN IMMUNODEFICIENCY VIRUS and is used in AIDS research.
A non-essential amino acid. In animals it is synthesized from PHENYLALANINE. It is also the precursor of EPINEPHRINE; THYROID HORMONES; and melanin.
Proteins obtained from the species SACCHAROMYCES CEREVISIAE. The function of specific proteins from this organism are the subject of intense scientific interest and have been used to derive basic understanding of the functioning similar proteins in higher eukaryotes.
In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.
Complexes of RNA-binding proteins with ribonucleic acids (RNA).
Theoretical representations that simulate the behavior or activity of biological processes or diseases. For disease models in living animals, DISEASE MODELS, ANIMAL is available. Biological models include the use of mathematical equations, computers, and other electronic equipment.
Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme.
A member of the p300-CBP transcription factors that was originally identified as a binding partner for ADENOVIRUS E1A PROTEINS.
A histone deacetylase subtype that is found along with HISTONE DEACETYLASE 2; RETINOBLASTOMA-BINDING PROTEIN 4; and RETINOBLASTOMA-BINDING PROTEIN 7 as core components of histone deacetylase complexes.
Single-stranded complementary DNA synthesized from an RNA template by the action of RNA-dependent DNA polymerase. cDNA (i.e., complementary DNA, not circular DNA, not C-DNA) is used in a variety of molecular cloning experiments as well as serving as a specific hybridization probe.
A chromatographic technique that utilizes the ability of biological molecules to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
Products of proto-oncogenes. Normally they do not have oncogenic or transforming properties, but are involved in the regulation or differentiation of cell growth. They often have protein kinase activity.
The complex formed by the binding of antigen and antibody molecules. The deposition of large antigen-antibody complexes leading to tissue damage causes IMMUNE COMPLEX DISEASES.
The part of a cell that contains the CYTOSOL and small structures excluding the CELL NUCLEUS; MITOCHONDRIA; and large VACUOLES. (Glick, Glossary of Biochemistry and Molecular Biology, 1990)
Proteins that control the CELL DIVISION CYCLE. This family of proteins includes a wide variety of classes, including CYCLIN-DEPENDENT KINASES, mitogen-activated kinases, CYCLINS, and PHOSPHOPROTEIN PHOSPHATASES as well as their putative substrates such as chromatin-associated proteins, CYTOSKELETAL PROTEINS, and TRANSCRIPTION FACTORS.
The determination of the pattern of genes expressed at the level of GENETIC TRANSCRIPTION, under specific circumstances or in a specific cell.
Techniques used to demonstrate or measure an immune response, and to identify or measure antigens using antibodies.
Reagents with two reactive groups, usually at opposite ends of the molecule, that are capable of reacting with and thereby forming bridges between side chains of amino acids in proteins; the locations of naturally reactive areas within proteins can thereby be identified; may also be used for other macromolecules, like glycoproteins, nucleic acids, or other.
Endogenous tissue constituents that have the ability to interact with AUTOANTIBODIES and cause an immune response.
Elements of limited time intervals, contributing to particular results or situations.
Proteins whose abnormal expression (gain or loss) are associated with the development, growth, or progression of NEOPLASMS. Some neoplasm proteins are tumor antigens (ANTIGENS, NEOPLASM), i.e. they induce an immune reaction to their tumor. Many neoplasm proteins have been characterized and are used as tumor markers (BIOMARKERS, TUMOR) when they are detectable in cells and body fluids as monitors for the presence or growth of tumors. Abnormal expression of ONCOGENE PROTEINS is involved in neoplastic transformation, whereas the loss of expression of TUMOR SUPPRESSOR PROTEINS is involved with the loss of growth control and progression of the neoplasm.
A continuous cell line of high contact-inhibition established from NIH Swiss mouse embryo cultures. The cells are useful for DNA transfection and transformation studies. (From ATCC [Internet]. Virginia: American Type Culture Collection; c2002 [cited 2002 Sept 26]. Available from http://www.atcc.org/)
Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.
Microscopy of specimens stained with fluorescent dye (usually fluorescein isothiocyanate) or of naturally fluorescent materials, which emit light when exposed to ultraviolet or blue light. Immunofluorescence microscopy utilizes antibodies that are labeled with fluorescent dye.
The first nucleotide of a transcribed DNA sequence where RNA polymerase (DNA-DIRECTED RNA POLYMERASE) begins synthesizing the RNA transcript.
Any of the processes by which nuclear, cytoplasmic, or intercellular factors influence the differential control of gene action during the developmental stages of an organism.
All of the processes involved in increasing CELL NUMBER including CELL DIVISION.
Detection of RNA that has been electrophoretically separated and immobilized by blotting on nitrocellulose or other type of paper or nylon membrane followed by hybridization with labeled NUCLEIC ACID PROBES.
DNA locations with the consensus sequence CANNTG. ENHANCER ELEMENTS may contain multiple copies of this element. E-boxes play a regulatory role in the control of transcription. They bind with basic helix-loop-helix (bHLH) type TRANSCRIPTION FACTORS. Binding specificity is determined by the specific bHLH heterodimer or homodimer combination and by the specific nucleotides at the 3rd and 4th position of the E-box sequence.
A polynucleotide consisting essentially of chains with a repeating backbone of phosphate and ribose units to which nitrogenous bases are attached. RNA is unique among biological macromolecules in that it can encode genetic information, serve as an abundant structural component of cells, and also possesses catalytic activity. (Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
Members of the class of compounds composed of AMINO ACIDS joined together by peptide bonds between adjacent amino acids into linear, branched or cyclical structures. OLIGOPEPTIDES are composed of approximately 2-12 amino acids. Polypeptides are composed of approximately 13 or more amino acids. PROTEINS are linear polypeptides that are normally synthesized on RIBOSOMES.
The mechanisms effecting establishment, maintenance, and modification of that specific physical conformation of CHROMATIN determining the transcriptional accessibility or inaccessibility of the DNA.
A family of zinc finger transcription factors that share homology with Kruppel protein, Drosophila. They contain a highly conserved seven amino acid spacer sequence in between their ZINC FINGER MOTIFS.
Different forms of a protein that may be produced from different GENES, or from the same gene by ALTERNATIVE SPLICING.
Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.
Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction.
Compounds that inhibit HISTONE DEACETYLASES. This class of drugs may influence gene expression by increasing the level of acetylated HISTONES in specific CHROMATIN domains.
The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.
An essential amino acid. It is often added to animal feed.
Strains of mice in which certain GENES of their GENOMES have been disrupted, or "knocked-out". To produce knockouts, using RECOMBINANT DNA technology, the normal DNA sequence of the gene being studied is altered to prevent synthesis of a normal gene product. Cloned cells in which this DNA alteration is successful are then injected into mouse EMBRYOS to produce chimeric mice. The chimeric mice are then bred to yield a strain in which all the cells of the mouse contain the disrupted gene. Knockout mice are used as EXPERIMENTAL ANIMAL MODELS for diseases (DISEASE MODELS, ANIMAL) and to clarify the functions of the genes.
An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.
One of the mechanisms by which CELL DEATH occurs (compare with NECROSIS and AUTOPHAGOCYTOSIS). Apoptosis is the mechanism responsible for the physiological deletion of cells and appears to be intrinsically programmed. It is characterized by distinctive morphologic changes in the nucleus and cytoplasm, chromatin cleavage at regularly spaced sites, and the endonucleolytic cleavage of genomic DNA; (DNA FRAGMENTATION); at internucleosomal sites. This mode of cell death serves as a balance to mitosis in regulating the size of animal tissues and in mediating pathologic processes associated with tumor growth.
A fractionated cell extract that maintains a biological function. A subcellular fraction isolated by ultracentrifugation or other separation techniques must first be isolated so that a process can be studied free from all of the complex side reactions that occur in a cell. The cell-free system is therefore widely used in cell biology. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p166)
The process by which two molecules of the same chemical composition form a condensation product or polymer.
Antibodies that react with self-antigens (AUTOANTIGENS) of the organism that produced them.
Histochemical localization of immunoreactive substances using labeled antibodies as reagents.
Cis-acting DNA sequences which can increase transcription of genes. Enhancers can usually function in either orientation and at various distances from a promoter.
Nucleoproteins, which in contrast to HISTONES, are acid insoluble. They are involved in chromosomal functions; e.g. they bind selectively to DNA, stimulate transcription resulting in tissue-specific RNA synthesis and undergo specific changes in response to various hormones or phytomitogens.
A sulfur-containing essential L-amino acid that is important in many body functions.
A specificity protein transcription factor that regulates expression of a variety of genes including VASCULAR ENDOTHELIAL GROWTH FACTOR and CYCLIN-DEPENDENT KINASE INHIBITOR P27.
Macromolecular complexes formed from the association of defined protein subunits.
Ubiquitous, inducible, nuclear transcriptional activator that binds to enhancer elements in many different cell types and is activated by pathogenic stimuli. The NF-kappa B complex is a heterodimer composed of two DNA-binding subunits: NF-kappa B1 and relA.
Cell surface proteins that bind signalling molecules external to the cell with high affinity and convert this extracellular event into one or more intracellular signals that alter the behavior of the target cell (From Alberts, Molecular Biology of the Cell, 2nd ed, pp693-5). Cell surface receptors, unlike enzymes, do not chemically alter their ligands.
Protein kinases that catalyze the PHOSPHORYLATION of TYROSINE residues in proteins with ATP or other nucleotides as phosphate donors.
Structurally related forms of an enzyme. Each isoenzyme has the same mechanism and classification, but differs in its chemical, physical, or immunological characteristics.
Cellular DNA-binding proteins encoded by the c-myc genes. They are normally involved in nucleic acid metabolism and in mediating the cellular response to growth factors. Elevated and deregulated (constitutive) expression of c-myc proteins can cause tumorigenesis.
Nuclear phosphoprotein encoded by the p53 gene (GENES, P53) whose normal function is to control CELL PROLIFERATION and APOPTOSIS. A mutant or absent p53 protein has been found in LEUKEMIA; OSTEOSARCOMA; LUNG CANCER; and COLORECTAL CANCER.
A strain of albino rat used widely for experimental purposes because of its calmness and ease of handling. It was developed by the Sprague-Dawley Animal Company.
Proteins and peptides that are involved in SIGNAL TRANSDUCTION within the cell. Included here are peptides and proteins that regulate the activity of TRANSCRIPTION FACTORS and cellular processes in response to signals from CELL SURFACE RECEPTORS. Intracellular signaling peptide and proteins may be part of an enzymatic signaling cascade or act through binding to and modifying the action of other signaling factors.
Glycoproteins found on the membrane or surface of cells.
Eukaryotic cell line obtained in a quiescent or stationary phase which undergoes conversion to a state of unregulated growth in culture, resembling an in vitro tumor. It occurs spontaneously or through interaction with viruses, oncogenes, radiation, or drugs/chemicals.
Genetically engineered MUTAGENESIS at a specific site in the DNA molecule that introduces a base substitution, or an insertion or deletion.
A group of enzymes that catalyzes the phosphorylation of serine or threonine residues in proteins, with ATP or other nucleotides as phosphate donors.
Single chains of amino acids that are the units of multimeric PROTEINS. Multimeric proteins can be composed of identical or non-identical subunits. One or more monomeric subunits may compose a protomer which itself is a subunit structure of a larger assembly.
A broad category of carrier proteins that play a role in SIGNAL TRANSDUCTION. They generally contain several modular domains, each of which having its own binding activity, and act by forming complexes with other intracellular-signaling molecules. Signal-transducing adaptor proteins lack enzyme activity, however their activity can be modulated by other signal-transducing enzymes
A member of the p300-CBP transcription factor family that was initially identified as a binding partner for CAMP RESPONSE ELEMENT-BINDING PROTEIN. Mutations in CREB-binding protein are associated with RUBINSTEIN-TAYBI SYNDROME.
Technique involving the diffusion of antigen or antibody through a semisolid medium, usually agar or agarose gel, with the result being a precipitin reaction.
Areas of increased density of the dinucleotide sequence cytosine--phosphate diester--guanine. They form stretches of DNA several hundred to several thousand base pairs long. In humans there are about 45,000 CpG islands, mostly found at the 5' ends of genes. They are unmethylated except for those on the inactive X chromosome and some associated with imprinted genes.
Components of a cell produced by various separation techniques which, though they disrupt the delicate anatomy of a cell, preserve the structure and physiology of its functioning constituents for biochemical and ultrastructural analysis. (From Alberts et al., Molecular Biology of the Cell, 2d ed, p163)
Nucleotide sequences of a gene that are involved in the regulation of GENETIC TRANSCRIPTION.
The region of DNA which borders the 5' end of a transcription unit and where a variety of regulatory sequences are located.
The chemical or biochemical addition of carbohydrate or glycosyl groups to other chemicals, especially peptides or proteins. Glycosyl transferases are used in this biochemical reaction.
Adherence of cells to surfaces or to other cells.
CELL LINE derived from the ovary of the Chinese hamster, Cricetulus griseus (CRICETULUS). The species is a favorite for cytogenetic studies because of its small chromosome number. The cell line has provided model systems for the study of genetic alterations in cultured mammalian cells.
One of the ESTROGEN RECEPTORS that has marked affinity for ESTRADIOL. Its expression and function differs from, and in some ways opposes, ESTROGEN RECEPTOR BETA.
A multistage process that includes cloning, physical mapping, subcloning, determination of the DNA SEQUENCE, and information analysis.
An ERYTHROLEUKEMIA cell line derived from a CHRONIC MYELOID LEUKEMIA patient in BLAST CRISIS.
Highly conserved nuclear RNA-protein complexes that function in RNA processing in the nucleus, including pre-mRNA splicing and pre-mRNA 3'-end processing in the nucleoplasm, and pre-rRNA processing in the nucleolus (see RIBONUCLEOPROTEINS, SMALL NUCLEOLAR).
A human liver tumor cell line used to study a variety of liver-specific metabolic functions.
A transferase that catalyzes the addition of aliphatic, aromatic, or heterocyclic FREE RADICALS as well as EPOXIDES and arene oxides to GLUTATHIONE. Addition takes place at the SULFUR. It also catalyzes the reduction of polyol nitrate by glutathione to polyol and nitrite.
The complex series of phenomena, occurring between the end of one CELL DIVISION and the end of the next, by which cellular material is duplicated and then divided between two daughter cells. The cell cycle includes INTERPHASE, which includes G0 PHASE; G1 PHASE; S PHASE; and G2 PHASE, and CELL DIVISION PHASE.
A family of DNA-binding transcription factors that contain a basic HELIX-LOOP-HELIX MOTIF.
An E2F transcription factor that interacts directly with RETINOBLASTOMA PROTEIN and CYCLIN A and activates GENETIC TRANSCRIPTION required for CELL CYCLE entry and DNA synthesis. E2F1 is involved in DNA REPAIR and APOPTOSIS.
Cells that line the inner and outer surfaces of the body by forming cellular layers (EPITHELIUM) or masses. Epithelial cells lining the SKIN; the MOUTH; the NOSE; and the ANAL CANAL derive from ectoderm; those lining the RESPIRATORY SYSTEM and the DIGESTIVE SYSTEM derive from endoderm; others (CARDIOVASCULAR SYSTEM and LYMPHATIC SYSTEM) derive from mesoderm. Epithelial cells can be classified mainly by cell shape and function into squamous, glandular and transitional epithelial cells.
A protein that has been shown to function as a calcium-regulated transcription factor as well as a substrate for depolarization-activated CALCIUM-CALMODULIN-DEPENDENT PROTEIN KINASES. This protein functions to integrate both calcium and cAMP signals.
The processes triggered by interactions of ANTIBODIES with their ANTIGENS.
An enzyme that catalyzes the methylation of the epsilon-amino group of lysine residues in proteins to yield epsilon mono-, di-, and trimethyllysine. EC 2.1.1.43.
Body organ that filters blood for the secretion of URINE and that regulates ion concentrations.
A subunit of NF-kappa B that is primarily responsible for its transactivation function. It contains a C-terminal transactivation domain and an N-terminal domain with homology to PROTO-ONCOGENE PROTEINS C-REL.
Deletion of sequences of nucleic acids from the genetic material of an individual.
A multiribosomal structure representing a linear array of RIBOSOMES held together by messenger RNA; (RNA, MESSENGER); They represent the active complexes in cellular protein synthesis and are able to incorporate amino acids into polypeptides both in vivo and in vitro. (From Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed)
Substances elaborated by viruses that have antigenic activity.
A light microscopic technique in which only a small spot is illuminated and observed at a time. An image is constructed through point-by-point scanning of the field in this manner. Light sources may be conventional or laser, and fluorescence or transmitted observations are possible.
An N-acetylglycosamine containing antiviral antibiotic obtained from Streptomyces lysosuperificus. It is also active against some bacteria and fungi, because it inhibits the glucosylation of proteins. Tunicamycin is used as tool in the study of microbial biosynthetic mechanisms.
A multiprotein complex composed of the products of c-jun and c-fos proto-oncogenes. These proteins must dimerize in order to bind to the AP-1 recognition site, also known as the TPA-responsive element (TRE). AP-1 controls both basal and inducible transcription of several genes.
A large superfamily of transcription factors that contain a region rich in BASIC AMINO ACID residues followed by a LEUCINE ZIPPER domain.

Elevated DNA double strand breaks and apoptosis in the CNS of scid mutant mice. (1/8836)

Genetic approaches have provided evidence that DNA end-joining problems serve an essential role in neuronal survival during development of mammalian embryos. In the present study, we tested whether the DNA repair enzyme, DNA dependent protein kinase, plays an important role in the survival of cerebral cortical neurons in mice. DNA-PK is comprised of a DNA-binding subunit called Ku and a catalytic subunit called DNA-PKcs. In mice with the scid mutation, DNA-PKcs is truncated near the kinase domain, which causes loss of kinase activity. We compared the spatial and temporal aspects of neuronal cell death in scid versus isogenic wild-type embryos and found a significant increase in dying cells in scid mice, as assessed by nuclear changes, DNA fragmentation and caspase-3 activity. Additional biochemical and immunocytochemical studies indicated that of several DNA repair enzymes investigated, only PARP was increased in scid mice, possibly in response to elevated DNA strand breaks.  (+info)

Complexes between the LKB1 tumor suppressor, STRAD alpha/beta and MO25 alpha/beta are upstream kinases in the AMP-activated protein kinase cascade. (2/8836)

BACKGROUND: The AMP-activated protein kinase (AMPK) cascade is a sensor of cellular energy charge that acts as a 'metabolic master switch' and inhibits cell proliferation. Activation requires phosphorylation of Thr172 of AMPK within the activation loop by upstream kinases (AMPKKs) that have not been identified. Recently, we identified three related protein kinases acting upstream of the yeast homolog of AMPK. Although they do not have obvious mammalian homologs, they are related to LKB1, a tumor suppressor that is mutated in the human Peutz-Jeghers cancer syndrome. We recently showed that LKB1 exists as a complex with two accessory subunits, STRAD alpha/beta and MO25 alpha/beta. RESULTS: We report the following observations. First, two AMPKK activities purified from rat liver contain LKB1, STRAD alpha and MO25 alpha, and can be immunoprecipitated using anti-LKB1 antibodies. Second, both endogenous and recombinant complexes of LKB1, STRAD alpha/beta and MO25 alpha/beta activate AMPK via phosphorylation of Thr172. Third, catalytically active LKB1, STRAD alpha or STRAD beta and MO25 alpha or MO25 beta are required for full activity. Fourth, the AMPK-activating drugs AICA riboside and phenformin do not activate AMPK in HeLa cells (which lack LKB1), but activation can be restored by stably expressing wild-type, but not catalytically inactive, LKB1. Fifth, AICA riboside and phenformin fail to activate AMPK in immortalized fibroblasts from LKB1-knockout mouse embryos. CONCLUSIONS: These results provide the first description of a physiological substrate for the LKB1 tumor suppressor and suggest that it functions as an upstream regulator of AMPK. Our findings indicate that the tumors in Peutz-Jeghers syndrome could result from deficient activation of AMPK as a consequence of LKB1 inactivation.  (+info)

Biochemical characterization of DNA damage checkpoint complexes: clamp loader and clamp complexes with specificity for 5' recessed DNA. (3/8836)

The cellular pathways involved in maintaining genome stability halt cell cycle progression in the presence of DNA damage or incomplete replication. Proteins required for this pathway include Rad17, Rad9, Hus1, Rad1, and Rfc-2, Rfc-3, Rfc-4, and Rfc-5. The heteropentamer replication factor C (RFC) loads during DNA replication the homotrimer proliferating cell nuclear antigen (PCNA) polymerase clamp onto DNA. Sequence similarities suggest the biochemical functions of an RSR (Rad17-Rfc2-Rfc3-Rfc4-Rfc5) complex and an RHR heterotrimer (Rad1-Hus1-Rad9) may be similar to that of RFC and PCNA, respectively. RSR purified from human cells loads RHR onto DNA in an ATP-, replication protein A-, and DNA structure-dependent manner. Interestingly, RSR and RFC differed in their ATPase activities and displayed distinct DNA substrate specificities. RSR preferred DNA substrates possessing 5' recessed ends whereas RFC preferred 3' recessed end DNA substrates. Characterization of the biochemical loading reaction executed by the checkpoint clamp loader RSR suggests new insights into the mechanisms underlying recognition of damage-induced DNA structures and signaling to cell cycle controls. The observation that RSR loads its clamp onto a 5' recessed end supports a potential role for RHR and RSR in diverse DNA metabolism, such as stalled DNA replication forks, recombination-linked DNA repair, and telomere maintenance, among other processes.  (+info)

The Drosophila sterile-20 kinase slik controls cell proliferation and apoptosis during imaginal disc development. (4/8836)

Cell proliferation and programmed cell death are closely controlled during animal development. Proliferative stimuli generally also induce apoptosis, and anti-apoptotic factors are required to allow net cell proliferation. Genetic studies in Drosophila have led to identification of a number of genes that control both processes, providing new insights into the mechanisms that coordinate cell growth, proliferation, and death during development and that fail to do so in diseases of cell proliferation. We present evidence that the Drosophila Sterile-20 kinase Slik promotes cell proliferation and controls cell survival. At normal levels, Slik provides survival cues that prevent apoptosis. Cells deprived of Slik activity can grow, divide, and differentiate, but have an intrinsic survival defect and undergo apoptosis even under conditions in which they are not competing with normal cells for survival cues. Like some oncogenes, excess Slik activity stimulates cell proliferation, but this is compensated for by increased cell death. Tumor-like tissue overgrowth results when apoptosis is prevented. We present evidence that Slik acts via Raf, but not via the canonical ERK pathway. Activation of Raf can compensate for the lack of Slik and support cell survival, but activation of ERK cannot. We suggest that Slik mediates growth and survival cues to promote cell proliferation and control cell survival during Drosophila development.  (+info)

T cell receptor-independent basal signaling via Erk and Abl kinases suppresses RAG gene expression. (5/8836)

Signal transduction pathways guided by cellular receptors commonly exhibit low-level constitutive signaling in a continuous, ligand-independent manner. The dynamic equilibrium of positive and negative regulators establishes such a tonic signal. Ligand-independent signaling by the precursors of mature antigen receptors regulates development of B and T lymphocytes. Here we describe a basal signal that controls gene expression profiles in the Jurkat T cell line and mouse thymocytes. Using DNA microarrays and Northern blots to analyze unstimulated cells, we demonstrate that expression of a cluster of genes, including RAG-1 and RAG-2, is repressed by constitutive signals requiring the adapter molecules LAT and SLP-76. This TCR-like pathway results in constitutive low-level activity of Erk and Abl kinases. Inhibition of Abl by the drug STI-571 or inhibition of signaling events upstream of Erk increases RAG-1 expression. Our data suggest that physiologic gene expression programs depend upon tonic activity of signaling pathways independent of receptor ligation.  (+info)

Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. (6/8836)

VCY2 is a testis-specific protein that locates in a frequently deleted azoospermia factor c region on chromosome Yq. Although its genomic structure has been characterized, the function of VCY2 is still unknown. To gain insight regarding the likely function of VCY2, we investigated the proteins that interact with VCY2 using the yeast two-hybrid system. We identified a novel VCY2 interaction partner, named VCY2IP-1, that encodes an open reading frame of 1059 amino acids. The amino acid sequence of VCY2IP-1 shows 59.3% and 41.9% homology to two human microtubule-associated proteins (MAPs), MAP1B and MAP1A, respectively. VCY2IP-1 has an extensive homology to the N-terminus and C-terminus regions of MAP1B and MAP1A, placing it within a large family of MAPs. We mapped VCY2IP-1 to chromosome 19p13.11. The VCY2IP-1 gene spans 15 kilobases (kb) and consists of seven exons. Northern blot analysis identified a single, intense band of approximately 3.2-kb VCY2IP-1 transcript, predominantly expressed in human testis. In situ hybridization of human testicular sections showed the localization of VCY2IP-1 transcripts in germ cells, and reverse transcription-polymerase chain reaction analysis demonstrated the presence of VCY2 and VCY2IP-1 transcripts in human ejaculated spermatozoa. Our expression data support the involvement of VCY2 and VCY2IP-1 in spermatogenesis. Based on the high homology of VCY2IP-1 with MAPs, we propose the involvement of VCY2 in the cytoskeletal network via interaction with VCY2IP-1.  (+info)

Identification of dimeric and oligomeric complexes of the human oxytocin receptor by co-immunoprecipitation and bioluminescence resonance energy transfer. (7/8836)

The nonapeptide hormone oxytocin exerts many important biological functions, including uterine contractions during parturition and milk ejection during lactation. The manifold effects of oxytocin are mediated by a single oxytocin receptor (OTR) type, a member of the super-family of G-protein-coupled receptors. There is accumulating recent evidence that certain G-protein-coupled receptors exist in the form of oligomeric complexes. Here we demonstrate, using two different co-immunoprecipitation strategies as well as bioluminescence resonance energy transfer techniques, that the OTR is capable of forming oligomeric complexes in vivo and that these complexes exist at the cell surface membrane. The human OTR was N-terminally tagged with either a Myc or Flag epitope and transiently expressed in COS-7 cells. Cell lysates were immunoprecipitated using an anti-Flag antibody and analyzed by SDS-PAGE and Western blotting using an anti-Myc antibody, or vice versa. Either strategy provided evidence for the co-precipitation of Myc- or Flag-tagged OTR respectively. Biochemical characterization of OTR dimers showed that homodimer formation is not dependent on the establishment of disulfide bonds. The existence of OTR dimers and oligomers at the level of the cell surface was demonstrated by exposing intact living cells to an anti-Flag antibody and analyzing the immunoprecipitate by Western blotting with an anti-Myc antibody. This approach demonstrated furthermore that the presence of receptor oligomers at the cell surface is modulated by ligand in a time-dependent fashion. Finally, we obtained evidence that the OTR is forming oligomeric structures in intact living cells by observing the occurrence of bioluminescence resonance energy transfer in cells co-transfected with OTR constructs bearing at their C-terminus either a Renilla luciferase or the yellow fluorescent protein. Taken together, these data show that the OTR can form homodimers and oligomers in the cell model used and that these oligomers are present at the cell surface.  (+info)

Impaired renal clearance explains elevated troponin T fragments in hemodialysis patients. (8/8836)

BACKGROUND: Patients with severe renal dysfunction often have unexplained elevated serum concentrations of cardiac troponin T (cTnT). We investigated whether in vivo fragmentation of cTnT could explain these increases. METHODS AND RESULTS: cTnT, creatine kinase isoenzyme MB, and myoglobin serum concentrations were measured in all 63 dialysis patients of our in-hospital dialysis department. A highly sensitive immunoprecipitation assay, followed by electrophoresis and Western blotting, was used to extract and concentrate cTnT and its possible fragments from serum of these 63 hemodialysis patients. Although creatine kinase isoenzyme MB values excluded recent ischemic myocardial events in 55 of the 63 cases, cTnT fragments ranging in size from 8 to 25 kDa were present in the serum samples of all dialysis patients. CONCLUSIONS: cTnT is fragmented into molecules small enough to be cleared by the kidneys of healthy subjects. Impaired renal function causes accumulation of these cTnT fragments and is very likely the cause of the unexplained elevations of serum cTnT found in patients with severe renal failure.  (+info)

The Thermo Scientific Pierce c-Myc Tag IP/Co-IP Kit provides the affinity resin and other reagents necessary to easily perform immunoprecipitation (IP) or co-immunoprecipitation (co-IP) experiments using a c-Myc-tagged protein as the bait.The c-Myc peptide (EQKLISEEDL) has become a popular fusion ta
Co-IP and WB analysis of cohesin Rad21-Rad21 interaction. Logarithmically growing 293T or HeLa cells were transfected with appropriate Rad21 plasmids or empty
Capturem Protein A miniprep columns were used to perform fast, efficient, and specific immunoprecipitation and co-immunoprecipitation experiments, and shown to be compatible with a variety of immunoprecipitation buffers.
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How many cells needed for co-IP and mass spec - posted in Protein and Proteomics: How many cells do I need for a co-IP followed by mass spec to identify binding partners? I have a stable cell line expressing a particular protein with a FLAG-HA tag. I would like to immunoprecipitate this protein (using Sigmas FLAG-HA tandem affinity purification kit). I would then like to use mass spec to identify all the binding partners of my FLAG-HA tagged protein. Approximately how many cells will I n...
12 days embryo embryonic body between diaphragm region and neck cDNA RIKEN full-length enriched library clone:9430068A02 product:zinc finger RNA binding protein full insert sequence ...
Mus musculus B6-derived CD11 +ve dendritic cells cDNA RIKEN full-length enriched library clone:F730023J15 product:hypothetical Prokaryotic membrane lipoprotein lipid attachment site containing protein full insert sequence ...
Adult inner ear cDNA RIKEN full-length enriched library clone:F930001H16 product:weakly similar to NOLP protein (HRIHFB2255 protein) (10 days neonate cortex cDNA RIKEN full-length enriched library clone:A830089E07 product:weakly similar to NOLP ...
The MethylPath MeDIP-Seq Service from Active Motif Epigenetic Services combines methylated DNA immunoprecipitation and Next-Gen sequencing to enable you to study differentially methylated regions of DNA throughout the entire genome.
Prevalence studies have demonstrated a global distribution of equine hepacivirus (EqHV), a member of the family Flaviviridae. However, apart from a single case of vertical transmission, natural routes of EqHV transmission remain elusive. Many known flaviviruses are horizontally transmitted between hematophagous arthropods and vertebrate hosts. This study represents the first investigation of potential EqHV transmission by mosquitoes. More than 5000 mosquitoes were collected across Austria and analyzed for EqHV ribonucleic acid (RNA) by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Concurrently, 386 serum samples from horses in eastern Austria were analyzed for EqHV-specific antibodies by luciferase immunoprecipitation system (LIPS) and for EqHV RNA by RT-qPCR. Additionally, liver-specific biochemistry parameters were compared between EqHV RNA-positive horses and EqHV RNA-negative horses. Phylogenetic analysis was conducted in comparison to previously published sequences ...
Adult male testis cDNA RIKEN full-length enriched library clone:4930562C03 product:hypothetical S-adenosyl-L-methionine- dependent methyltransferases structure containing protein full insert sequence (Adult male olfactory brain cDNA RIKEN full- ...
Gels are used in immunoprecipitation techniques to stabilize the precipitate, enabling both the position and the area of the precipitate to be measured. The
Immunoprecipitation and Western blot analysis. a. Immunoprecipitation: lung extract from E15.5 wild-type embryos was used for immunoprecipitation using Smad2 an
Co-inmunoprecipitación (CoIP) y ensayos de pull-down son métodos de cerca relacionados para identificar las interacciones proteína-proteína estable. Estos...
could anyone of u suggest a good rabbit Isotype Control for co-ip experiments since my first antibody is raised in rabbit . i gave a search on all company websites, they dont have one which is compatible for western blots. can i use ELISA/FACS compatible abs for the same ...
An important tool for studying the regulation of synapses is a rapid and reliable means of separating synaptic and intracellular proteins. This unit presents a technique for analysis of brain tissue which relies on differential centrifugation to separate proteins present at synaptic sites from those found in intracellular cytoplasmic and vesicular pools. The method is efficient in that only small amounts of tissue, such as might be obtained from a small region of a rodent brain, are required. It is reproducible and, in conjunction with immunoblot or immunoprecipitation techniques, can produce reliable quantitative data. The protocol will be of interest to those conducting a variety of different studies related to the localization and trafficking of brain receptors and signaling molecules. Curr. Protoc. Neurosci. 42:1.16.1-1.16.16. © 2008 by John Wiley & Sons, Inc. ...
An important tool for studying the regulation of synapses is a rapid and reliable means of separating synaptic and intracellular proteins. This unit presents a technique for analysis of brain tissue which relies on differential centrifugation to separate proteins present at synaptic sites from those found in intracellular cytoplasmic and vesicular pools. The method is efficient in that only small amounts of tissue, such as might be obtained from a small region of a rodent brain, are required. It is reproducible and, in conjunction with immunoblot or immunoprecipitation techniques, can produce reliable quantitative data. The protocol will be of interest to those conducting a variety of different studies related to the localization and trafficking of brain receptors and signaling molecules. © 2008 by John Wiley & Sons, Inc ...
Catch & Release v2.0 High Throughput (HT) Immunoprecipitation Assay Kit- 96 well from Upstate,This kit allows for quick and reproducible immunoprecipitation (IP) by using a 96 well filter plate. The system is more reproducible than regular IPs, which are problematic with regards to washing the protein A/G agarose without disrupting the agarose bed. The binding of the antibody/antigen comple,biological,biology supply,biology supplies,biology product
Long noncoding RNAs (lncRNAs) have important roles in shaping chromatin by targeting chromatin-modifying enzymes to distinct genomic sites. This section covers two methods to analyze lncRNA-protein interactions. The RNA-protein pull-down assays use either bead-bound proteins to capture in vitro transcripts, or immobilized synthetic RNAs to bind proteins from cell lysates. In the RNA immunoprecipitation (RIP) assay, endogenous RNAs are co-immunoprecipitated with a protein of interest. Both the methods can be applied to material from proliferating and quiescent cells, thus providing insights into how lncRNA-protein interactions are altered between these two cellular states.. Schematic overview of the lncRNA-protein interaction assays described in this protocol. ...
An automated instrument that standardizes different Epigenetic Applications such as Chromatin Immunoprecipitation (ChIP), Methylated DNA Immunoprecipitation (MeDIP), Bisulfite Conversion..
Hyperphosphatemic familial tumoral calcinosis (HFTC)/hyperostosis-hyperphosphatemia syndrome (HHS) is an autosomal recessive disorder of ectopic calcification due to deficiency of or resistance to intact fibroblast growth factor 23 (iFGF23). Inactivating mutations in FGF23, N-acetylgalactosaminyltransferase 3 (GALNT3), or KLOTHO (KL) have been reported as causing HFTC/HHS. We present what we believe is the first identified case of autoimmune hyperphosphatemic tumoral calcinosis in an 8-year-old boy. In addition to the classical clinical and biochemical features of hyperphosphatemic tumoral calcinosis, the patient exhibited markedly elevated intact and C-terminal FGF23 levels, suggestive of FGF23 resistance. However, no mutations in FGF23, KL, or FGF receptor 1 (FGFR1) were identified. He subsequently developed type 1 diabetes mellitus, which raised the possibility of an autoimmune cause for hyperphosphatemic tumoral calcinosis. Luciferase immunoprecipitation systems revealed markedly elevated ...
Hyperphosphatemic familial tumoral calcinosis (HFTC)/hyperostosis-hyperphosphatemia syndrome (HHS) is an autosomal recessive disorder of ectopic calcification due to deficiency of or resistance to intact fibroblast growth factor 23 (iFGF23). Inactivating mutations in FGF23, N-acetylgalactosaminyltransferase 3 (GALNT3), or KLOTHO (KL) have been reported as causing HFTC/HHS. We present what we believe is the first identified case of autoimmune hyperphosphatemic tumoral calcinosis in an 8-year-old boy. In addition to the classical clinical and biochemical features of hyperphosphatemic tumoral calcinosis, the patient exhibited markedly elevated intact and C-terminal FGF23 levels, suggestive of FGF23 resistance. However, no mutations in FGF23, KL, or FGF receptor 1 (FGFR1) were identified. He subsequently developed type 1 diabetes mellitus, which raised the possibility of an autoimmune cause for hyperphosphatemic tumoral calcinosis. Luciferase immunoprecipitation systems revealed markedly elevated ...
The in vivo association of histone H1 with specific genes in Tetrahymena thermophila was studied by using a simplified cross-linking and immunoprecipitation technique. Four genes were analyzed whose activities vary in three different developmental states (logarithmic growth, starvation, and conjugation). Hybridization of the immunoprecipitated DNA to cloned probes showed an inverse correlation between the level of immunoprecipitation with H1 antiserum and transcriptional activity. This represents the first demonstration of an alteration in histone H1-DNA interaction associated with developmental changes in transcriptional activity. ...
Supplementary MaterialsFIGURE S1: The PfDis3-ADARcd reproducibly edits particular sites in through the IDC. the editing rate of recurrence in the transcripts with an increase of than one edit sites. Picture_1.JPEG (1.6M) GUID:?DA3700B1-A11E-4C9E-ACF2-4216D199F6B4 FIGURE S2: Reproducibility of PfDis3-RIP assay across developmental phases. Relationship of genic RIP indicators in feeling (s) transcripts and antisense (as) transcripts Rabbit Polyclonal to IRAK2 between natural replicates. Pearson relationship coefficients between natural replicates are shown at top remaining part. The inset Venn diagram displaying the overlap of PfDis3 focuses on determined by PfDis3-RIP between your two replicates. R, T, S shows Ring, Schizont and Trophozoite stage, respectively. Picture_2.jpg (1.3M) GUID:?3CE2FFB1-19A0-4EC3-8727-049FC475ADF8 FIGURE S3: Functions of PfDis3-TRIBE target genes through the IDC in strategies of RIP-seq, HITS-CLIP, or GoldCLIP because of the high history and complicated manipulation ...
Chromatrap® offers a one-of-a-kind solid state patented technology, which is characterized by unmatched sensitivity. This makes it possible for users to perform Chromatin immunoprecipitation (ChIP) assays using only 1000 cells per immunoprecipitation.
The Enzyme Collection contains over 550 mAbs that recognize catalytic domains or associated regulatory subunits in enyme complexes
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RNA binding proteins are fundamental components of crucial biological processes in all domains of life. The development of techniques to study RNA binding protein complexes is, therefore, broadly impactful. I employed multiple genome-wide techniques to investigate the RNA binding repertoire of the histone stem-loop binding protein (SLBP) - namely, RIP-chip, RIP-seq and HITS-CLIP. These genome-wide approaches provide a comprehensive view of the ribonucleic acid components of the histone stem-loop mRNP complex. Also, the HITS-CLIP method reveals sites of molecular contact. All three techniques corroborated SLBPs specificity for histone mRNA. I developed a bioinformatic HITS-CLIP analysis pipeline that includes: (1) a clustering method for grouping genes by sequence coverage patterns, (2) a method for inference of nuclease cleavage sites to map RNP boundaries and (3) crosslink-induced mutation mapping to identify sites of molecular contact. I cross-validated my findings with crystallographic data ...
IL-8/CXCL8 Immunoprecipitation (IP) Kit are used for Immunoprecipitation of IL-8/CXCL8 protein which expressed in vitro expression systems.
Hi, I am trying to do a co-IP experiment on a HA tagged protein, and I eluted the protein with SDS buffer. However, the protein level is very low that after I ran the eluate on a gel followed by silver staining, I couldnt see any band. So now I am thinking combining all the eluate from multiple experiments. Is there a way that I can concentrate all these proteins? Will TCA precipitation work in the presence of SDS? Or is there any kind of concentrator that I can use? Thank you very much ...
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The Enzyme Collection contains over 550 mAbs that recognize catalytic domains or associated regulatory subunits in enyme complexes
Background. DNA methylation, referring to the reversible methylation of the 5 position of cytosine by methyltransferases, is a major epigenetic modification in multicellular organisms. In mammals, this modification primarily occurs at CpG sites, which in turn tend to cluster in regions called CpG islands. There is a small fraction of CpG islands that can overlap or be in close proximity to promoter regions of transcription start sites. The modification may also occur at other sites, but methylation at either of these sites can repress gene expression by either interfering with the binding of transcription factors or modifying chromatin structure to a repressive state.. Disease condition studies have largely fueled the effort in understanding the role of DNA methylation. Currently, the major research interest lies in investigating disease conditions such as cancer to identify regions of the DNA that has undergone extensive methylation changes. The genes contained in these regions are of ...
To find out whether the AGO-miRNA complex is more sensitive to the accessibility of a particular region inside the seed match, we analyze in detail the accessibility of a wide set of miRNA binding sites validated by PAR-CLIP and HITS-CLIP experiments. Our analysis reveals that nucleotides at the 3′-end of bound seed matches are significantly more accessible than nucleotides at the 5′-end as well as nucleotides at any positions in the unbound seed matches. We show that the accessibility of a single nucleotide at the 3′-end is more effective than the accessibility of several nucleotides at the 5′-end in discriminating between functional and nonfunctional binding sites. Analysis of mRNA and protein fold changes induced by miRNA overexpression demonstrates that genes with accessible nucleation regions at the 3′-end are down-regulated more strongly than genes whose accessible nucleation regions are located elsewhere within the seed match. We also observed an increase in the precision of the ...
View mouse Cavin3 Chr7:105480083-105482300 with: phenotypes, sequences, polymorphisms, proteins, references, function, expression
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
Hepatoma-derived growth factor (HDGF) is a growth factor related to normal development and tumorigenesis; however, the mechanism of its mitogenic and angiogenic activity still remains unknown. Analysis of the HDGF interactome could be important for understanding its function and integrative mechanisms, because knowledge about HDGF interactors is very limited. In this study, through streptavidin-binding peptide (SBP) and Flag tag-based tandem affinity purification (SBP/Flag-TAP) coupled with LC-MS/MS, 106 proteins were shown to form complexes with HDGF. RNAs were also found in the HDGF complex through the SBP-tag based RNA co-immunoprecipitation (SBP-RIP) assay. Some of these interactions were confirmed by Co-IP and RT-PCR. We then found that the HATH domain was essential for HDGF interactions including protein-protein and protein-RNA interactions, and that in the absence of the HATH domain, NO-HATH could not form complex. The interactome suggests that HDGF is a multifunctional protein and participates
In 2012, Roux et al. published a nice paper, that received no less than four article recommendations from F1000 researchers. The paper described a method for tracking the interaction partners a protein has had within a cell (a history of its interacting partners). The method, called BioID, is based on proximity-dependent biotinylation of proteins by a promiscuous biotin ligase mutant BirA (R118G), which is fused to your protein of interest. After an overnight incubation with biotin, cells can be subjected to harsh lysis and biotinylated proteins can be isolated and identified by mass spectroscopy to determine the proteins that had come into contact with the chimeric BirA (R118G) protein. This method is a bit different from standard co-IP or pull-down experiments, because it allows one to identify proteins who interact transiently or weakly with the protein of interest. Also, due to the strong biotin-avidin binding affinity, harsh washes can greatly reduce background protein binding. [Read ...
Research literature citing and demonstrating the increasing use of Dynabeads magnetic beads for immunoprecipitation (IP) experiments.
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They are also used in immunoprecipitation to separate proteins and anything bound to them (co-immunoprecipitation) from other ... Williams NE (2000). Immunoprecipitation procedures. Methods in Cell Biology. 62. San Diego, CA : Academic Press. pp. 449-53. ...
Chromatin Immunoprecipitation. Methods in Molecular Biology. 1689. pp. 83-101. doi:10.1007/978-1-4939-7380-4_8. ISBN 978-1-4939 ...
In 1984, John T. Lis innovated the Chromatin immunoprecipitation technique. In 1993, the Nuclear Ligation Assay was published, ... Chromatin Immunoprecipitation Assays. review. Methods in Molecular Biology. 567. pp. 171-88. doi:10.1007/978-1-60327-414-2_12. ...
Seale B, Lam C, Rackus DG, Chamberlain MD, Liu C, Wheeler AR (October 2016). "Digital Microfluidics for Immunoprecipitation". ... as proof of concept work for immunoprecipitation using digital microfluidics.5 DNA extraction from a whole blood sample has ...
Hempelmann E, Wilson RJ (1982). "Immunoprecipitation of malarial enzymes". Protozoology. 29: 637. Blood photomicrographs. ...
SDS-PAGE Hempelmann, E.; Putfarken, B.; Rangachari, K.; Wilson, R.J.M. (1986). "Immunoprecipitation of malarial acid ...
Ji, Hong (2010-08-01). "Lysis of Cultured Cells for Immunoprecipitation". Cold Spring Harbor Protocols. 2010 (8): pdb.prot5466 ... Sefton, Bartholomew M. (2001-01-01). "Labeling Cultured Cells with32Piand Preparing Cell Lysates for Immunoprecipitation". ... Nonidet P-40 or Triton X-100 50 mM Tris-Cl Adjust pH to 7.4 RIPA buffer is a commonly used lysis buffer for immunoprecipitation ... Labeling Cultured Cells with 32Pi and Preparing Cell Lysates for Immunoprecipitation. Current Protocols in Molecular Biology. ...
The liquid phase ligand binding assay of Immunoprecipitation (IP) is a method that is used to purify or enrich a specific ... "Immunoprecipitation (IP) technical guide and protocols" (PDF). Thermo Fisher Scientific Inc. Archived from the original (PDF) ...
This particular procedure is known as immunoprecipitation. Immunoprecipitation is quite capable of generating an extremely ...
immunoprecipitation ChIP-sequencing DNA sequencing Ginno, PA; Lott, PL; Christensen, HC; Korf, I; Chédin, F (30 March 2012). "R ... S9.6 mAb was first created and characterized in 1986 and is currently used for the selective immunoprecipitation of R-loops. ... Due to the absence of another antibody-based method for R-loop immunoprecipitation, validation of DRIP-seq results is difficult ... Since then, it was used in diverse immunoprecipitation methods for R-loop characterization. The concept behind DRIP-seq is ...
Jenkins CW, Xiong Y (1996). "Immunoprecipitation and Immunoblotting in Cell Cycle Studies". Cell Cycle - Materials and Methods ...
"Isolating human transcription factor targets by coupling chromatin immunoprecipitation". Genes Dev. 16 (2): 235-244. doi: ...
"Mass spectrometry-based immuno-precipitation proteomics - the user's guide". Proteomics. 11 (6): 1153-9. doi:10.1002/pmic. ...
This method relies upon immunoprecipitation of RNA polymerase II. There are a number of issues with immunoprecipitation, ... including non-specific binding interactions which may result in the immunoprecipitation of off-target RNA molecules. The ...
The aforementioned mRNA was isolated using antibody based immunoprecipitation. The resulting cDNA library was subsequently ...
Immunoprecipitation assays show that INSL3 does bind to NR4A1; however, much is still not known about INSL3 regulation and the ...
"Use of protein biotinylation in vivo for chromatin immunoprecipitation". Analytical Biochemistry. 325 (1): 68-76. doi:10.1016/j ...
MeDIP (methylated DNA immunoprecipitation) is an experimental technique used to assess DNA methylation levels by using an ... A Bayesian deconvolution strategy for immunoprecipitation-based DNA methylome analysis. Nature Biotechnology 26, 779-85 (2008 ... is a statistical tool for analysing methylated DNA immunoprecipitation (MeDIP) profiles. It can be applied to large datasets ...
This can be determined using a chromatin immunoprecipitation (ChIP) assay. DNA-nucleosome interactions are characterized by two ...
Quantitative immunoprecipitation combined with knock-down (QUICK) relies on co-immunoprecipitation, quantitative mass ... Co-immunoprecipitation is considered[citation needed] to be the gold standard assay for protein-protein interactions, ... Thus, it has the same high confidence as co-immunoprecipitation. However, this method also depends on the availability of ... A note of caution also is that immunoprecipitation experiments reveal direct and indirect interactions. Thus, positive results ...
Weinmann AS, Bartley SM, Zhang T, Zhang MQ, Farnham PJ (October 2001). "Use of chromatin immunoprecipitation to clone novel E2F ...
The interaction was detected by immunoprecipitation by Abdelmohsen et al., 2009. ELAVL1 is involved in regulating gene ...
"Purification of galactokinase mRNA from Saccharomyces cerevisiae by indirect immunoprecipitation". The Journal of Biological ...
α-dystroglycan was shown to interact with pikachurin through immunoprecipitation. Dystroglycan ligand with other proteins is ...
This procedure is essentially an immunoprecipitation (IP) of the protein. This can be done either by using a tagged protein ... ChIP-on-chip (also known as ChIP-chip) is a technology that combines chromatin immunoprecipitation ('ChIP') with DNA microarray ... Introduced in 2007, ChIP sequencing (ChIP-seq) is a technology that uses chromatin immunoprecipitation to crosslink the ... Aparicio, O; Geisberg, JV; Struhl, K (2004). Chromatin immunoprecipitation for determining the association of proteins with ...
Immunoprecipitation kinase assays revealed that cyclin C has Rb kinase activity. Furthermore, unlike cyclins D and E, cyclin ... Finally, co-immunoprecipitation assays revealed that cyclin-dependent kinase 3 (cdk3) promotes G0 exit by forming a complex ...
Sengupta M, Jain V, Wilkinson BJ, Jayaswal RK (June 2012). "Chromatin immunoprecipitation identifies genes under direct VraSR ...
The immunoprecipitation step also allows for the removal of non-specific binding sites. The fourth step is DNA recovery and ... The third step is called chromatin immunoprecipitation, which is what ChIP is short for. The ChIP process enhance specific ... Sono-Seq, identical to ChIP-Seq but skipping the immunoprecipitation step. HITS-CLIP (also called CLIP-Seq), for finding ... ChIP-seq combines chromatin immunoprecipitation (ChIP) with massively parallel DNA sequencing to identify the binding sites of ...
Protein complex immunoprecipitation (Co-IP)[edit]. Immunoprecipitation of intact protein complexes (i.e. antigen along with any ... RNP Immunoprecipitation (RIP)[edit]. Similar to chromatin immunoprecipitation (ChIP) outlined above, but rather than targeting ... Analysis of Proteins Using Immunoprecipitation at ufl.edu. *Immunoprecipitation at the US National Library of Medicine Medical ... Immunoprecipitation. Current Protocols in Molecular Biology. 10.16.1-10.16.29. *^ a b c Rosenberg, Ian (2005). Protein analysis ...
Anderson N.G. (1998) Co-Immunoprecipitation. In: Clegg R.A. (eds) Protein Targeting Protocols. Methods in Molecular Biology™, ... This chapter aims to describe one such technique, that of co-immunoprecipitation. ...
... Peter Corish pcor at bioch.ox.ac.uk Tue Jul 9 09:12:33 EST 1996 *Previous message: Confocal Newsgroup ... Has anybody had any experience (good or bad) using the advertised Clontech polyclonal GFP antibody for immunoprecipitations or ...
This makes it possible for users to perform Chromatin immunoprecipitation (ChIP) assays using only 1000 cells per ... Chromatin immunoprecipitation. The Chromatrap® ChIP-seq Protein A kit (Cat no 500189) was used for ChIP in accordance with the ... Efficient Extraction and Immunoprecipitation of Chromatin from Complex Formalin Fixed Paraffin Embedded Tissue. ... Slurries were prepared for immunoprecipitation using 100 µl chromatin stock from each extraction, depending on the number of ...
This up-to-date volume includes protocols that illustrate the broad use of chromatin immunoprecipitation (ChIP) and ChIP- ... Chromatin Immunoprecipitation. Methods and Protocols. Editors: Visa, Neus, Jordán-Pla, Antonio (Eds.) ... Chromatin Immunoprecipitation from Mouse Embryonic Tissue or Adherent Cells in Culture, Followed by Next-Generation Sequencing ... Includes cutting-edge techniques for the study of chromatin immunoprecipitation. *Provides step-by-step detail essential for ...
4: Co-immunoprecipitation of LONP1 and mtHSP70. , Nature Communications. Fig. 4: Co-immunoprecipitation of LONP1 and mtHSP70.. ...
... Ian A. York iayork at panix.com Mon Sep 13 07:31:12 EST 1999 *Previous message: Immunoprecipitations?? ...
Chromatin immunoprecipitation (ChIP) is a type of immunoprecipitation experimental technique used to investigate the ... Chromatin+immunoprecipitation at the US National Library of Medicine Medical Subject Headings (MeSH) EpigenomeNOE.com Chromatin ... Following lysis of cross-linked cells and immunoprecipitation of bacterial RNA polymerase, DNA associated with enriched RNA ... These reports are considered the pioneering studies in the field of chromatin immunoprecipitation. XChIP was further modified ...
Chromatin Immunoprecipitation Sequencing (ChIP Seq) News and Research. RSS ChIP-seq, or chromatin immunoprecipitation and ...
Discover more on advanced immunoprecipitation (IP) with this on-demand webinar. Learn how to identify protein interactions ... Next, I will discuss endogenous versus overexpressed protein immunoprecipitation. Immunoprecipitation of endogenous proteins ... and it will tell you how efficient your immunoprecipitation was. The third lane is the immunoprecipitation, so the outcome of ... Kits for the immunoprecipitation of methylated DNA are also available. To get an overview of all kits, please go to www.abcam. ...
113 Pages Report][Single User License:: US $5650] Immunoprecipitation Market categories the global market by Product (Kit, ... 6 Immunoprecipitation Market, By Type (Page No. - 34). 6.1 Introduction 6.2 Individual Immunoprecipitation 6.2.1 Increasing ... Table 4 Co-Immunoprecipitation Market, By Region, 2016 2024 (USD Million). Table 5 North America: Co-Immunoprecipitation Market ... Table 8 Rna Immunoprecipitation Market, By Region, 2016 2024 (USD Million). Table 9 North America: Rna Immunoprecipitation ...
Immunoprecipitation * Immunoprecipitation with Resins and Microcolumns * ChIP, RIP, and Protein-Nucleic Acid Pull-Down Products ... Epitope tags are seldom used for bulk purification but are most often used for immunoprecipitation or co-immunoprecipitation ( ... Epitope tags are seldom used for bulk purification but are most often used for immunoprecipitation or co-immunoprecipitation ( ... immunoprecipitation, IP) and protein complexes (co-immunoprecipitation, co-IP). ...
Immunoprecipitation Protocol Utilizing Magnetic Separation (For Analysis By Western Blot): easy to follow directions describing ... Proceed to immunoprecipitation section.. Immunoprecipitation. IMPORTANT: Appropriate isotype controls are highly recommended in ... Immunoprecipitation Protocol Utilizing Magnetic Separation (For Analysis By Western Immunoblotting). Immunoprecipitation ... C. Immunoprecipitation. Cell Lysate Pre-Clearing (Highly Recommended). A cell lysate pre-clearing step is highly recommended to ...
Chromatin immunoprecipitation (ChIP) is a powerful research technique used to identify and analyze protein-DNA interactions ...
The RNA immunoprecipitation (RIP) technique has been used to identify a large number of Hfq-associated RNA species, and can be ... Here, endogenously formed Hfq-RNA complexes of Y. pestis were captured with immunoprecipitation with a monoclonal anti-Flag ... Han Y. (2018) Purification of Hfq-Associated RNAs with RNA Immunoprecipitation (RIP). In: Yang R. (eds) Yersinia Pestis ...
A complete solution for Chromatin Immunoprecipitation including columns and reagents for DNA purification. ... The Imprint Chromatin Immunoprecipitation Kit combines speed and convenience with performance. ... Successful chromatin immunoprecipitation requires an effective, specific, and high quality antibody. Not all antibodies work in ... Sigma combines speed and convenience with performance in the Imprint Chromatin Immunoprecipitation Kit (CHP1). The Imprint ChIP ...
Home Protocols Immunoprecipitation using Protein A/G Magnetic Beads Immunoprecipitation using Protein A/G Magnetic Beads. ... Immunoprecipitation (1) This step pre-clears crude cell extract of proteins which can bind non-specifically to the beads. In a ... 2) Lyse the cells with 0.5 ml cold Immunoprecipitation Buffer (150 mM NaCl, 10 mM Tris-HCl (pH 7.4), 1 mM EDTA, 1 mM EGTA (pH ... Alternatively, Protein G Magnetic Beads (NEB #S1430S) can be used for immunoprecipitations with monoclonal antibodies. (3) Add ...
... immunoprecipitation HA-tagged fusion protein from cell lysates 2.5-4μg/test,indirect immunofluorescence HA-tagged fusion ... Anti-HA antibody is suitable for use in immunoprecipitation and western blot. It is also suitable for indirect ... immunoprecipitation (IP): 2.5-4 μg/test using HA-tagged fusion protein from cell lysates ... immunoprecipitation HA-tagged fusion protein from cell lysates 2.5-4μg/test, Ha Antibody Sigma, Ha Antibody ...
Immunoprecipitation analysis. A standard set of images defining the immunoprecipitation patterns of reactivity is shown below ... Immunoprecipitation analysis Serum samples were tested using the standard immunoprecipitation protocol (Reeves WH,et al., 2006 ... Immunoprecipitation analysis Testing of IgG autoantibodies to human cellular antigens was performed on samples that showed a 3+ ... IP: immunoprecipitation; Y: Identity can be confirmed; (Y) identity suggested but requires RNA-IP to confirm; ND: not done ...
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This video demonstrates the protocol for methylated DNA immunoprecipitation (MeDIP). MeDIP is a two day procedure that ... Methylated DNA Immunoprecipitation. Kelsie L. Thu1,2,3, Emily A. Vucic1,3,4, Jennifer Y. Kennett1,4, Cameron Heryet5, Carolyn J ... Methylated DNA immunoprecipitation (MeDIP) is an efficient technique for the extraction of methylated DNA from a sample of ... A sample of as little as 200 ng of DNA is sufficient for the antibody, or immunoprecipitation (IP), reaction. DNA is sonicated ...
Tools for Chromatin Immunoprecipitation Description ChIP Kit EZ-ChIP Kit Acetyl-Histone H3 Immunoprecipitation (ChIP) Assay Kit ... G Chromatin Immunoprecipitation Kit Magna ChIP™ A/G Chromatin Immunoprecipitation Kit EZ-Magna ChIP™ A/G Chromatin ... A Chromatin Immunoprecipitation Kit Magna ChIP™ G Chromatin Immunoprecipitation Kit EZ-Magna ChIP™ A Chromatin ... Universal Chromatin Immunoprecipitation DNA Microarray Quad Kit Magna ChIP-Seq Chromatin Immunoprecipitation and Next ...
... can be used to study protein DNA complexes. Visit this page to learn more about how to optimize ... Immunoprecipitation. Immunoprecipitation of chromatin-bound protein can be performed with Protein A, G, or A/G magnetic or ... Chromatin immunoprecipitation (ChIP) can be used as a tool to study protein:DNA complexes and identify protein-binding sites in ... Chromatin Immunoprecipitation is a powerful tool to study protein:DNA complexes and can be used to map transcription factor ...
Biological context of Immunoprecipitation. *Based on a co-immunoprecipitation of major histocompatibility complex (MHC) class I ... Disease relevance of Immunoprecipitation. *This protein was shown by immunoprecipitation to have antigenic determinants of MuLV ... Northern blot, immunoprecipitation, and flow cytometry analyses revealed that neutrophils did not express Fc epsilon RII/CD23, ... Co-immunoprecipitation studies demonstrate that RGS10 associates specifically with the activated forms of two related G-protein ...
Analysis of Histone Modifications in Rodent Pancreatic Islets by Native Chromatin Immunoprecipitation.. Sandovici I1,2, ...
Measurement of protein-DNA interactions in vivo by chromatin immunoprecipitation.. Im H1, Grass JA, Johnson KD, Boyer ME, Wu J ... The chromatin is sonicated to generate small fragments, and an immunoprecipitation is conducted with an antibody against the ... These seemingly complex issues can be directly addressed by a powerful methodology termed the chromatin immunoprecipitation ( ...
... combining chromatin immunoprecipitation (ChIP) and massively parallel sequencing to identify mammalian DNA sequences bound by ... Genome-wide profiles of STAT1 DNA association using chromatin immunoprecipitation and massively parallel sequencing. *Gordon ... We developed a method, ChIP-sequencing (ChIP-seq), combining chromatin immunoprecipitation (ChIP) and massively parallel ... Genome-wide profiles of STAT1 DNA association using chromatin immunoprecipitation and massively parallel sequencing. Nat ...
Immunoprecipitation Troubleshooting - Immunoprecipitation (Nov/02/2010 ). Hi all,. I need some help.We assume there is an ...
... a new article that describes a detailed genome wide comparison of native and crosslinked chromatin immunoprecipitation followe ... More From BioPortfolio on "Comparative Study of Native & Crosslinked Immunoprecipitation". *Related Companies *Related Events * ... genome wide comparison of native and crosslinked chromatin immunoprecipitation followed by next generation sequencing (ChIP-seq ... Comparative Study of Native & Crosslinked Immunoprecipitation. 14:14 EST 16 Jan 2018 , Lab Bulletin ...
  • Immunoprecipitation of intact protein complexes (i.e. antigen along with any proteins or ligands that are bound to it) is known as co-immunoprecipitation (Co-IP). (wikipedia.org)
  • As successive rounds of targeting and immunoprecipitations take place, the number of identified proteins may continue to grow. (wikipedia.org)
  • Immunoprecipitation of proteins can elucidate the molecular importance protein-protein interactions play in signal transduction. (abcam.com)
  • Chromatin immunoprecipitation (ChIP) is a type of immunoprecipitation experimental technique used to investigate the interaction between proteins and DNA in the cell. (wikipedia.org)
  • Because Dynabeads magnetic beads offer you the best balance of capacity/yield, reproducibility, purity, and cost for smaller-scale isolation of specific proteins (e.g., immunoprecipitation, IP) and protein complexes ( co-immunoprecipitation, co-IP ). (thermofisher.com)
  • Magnetic beads have become the gold standard for immunoprecipitation and pull-down assays because they offer a faster, easier, and more efficient way of pulling down proteins of interest than traditional Sepharose agarose or other agarose-based resins. (thermofisher.com)
  • One of the most commonly used methods to isolate proteins from their biological sources is immunoprecipitation (IP). (mt.com)
  • Immunoprecipitation (IP) is one of the most widely used techniques to isolate proteins and other biomolecules from complex mixture. (antibodies-online.com)
  • Identification of Akt-interacting proteins has been customarily achieved by co-immunoprecipitation combined with western blot and/or MS analysis. (nih.gov)
  • The kit is also optimized to give superior results when performing chromatin immunoprecipitation using antibodies against low abundance proteins, such as transcription factors , or antibodies with sub-optimal binding affinities. (activemotif.com)
  • Following immunoprecipitation, the DNA cross-links are reversed, the proteins are removed by Proteinase K and the DNA is recovered and purified. (activemotif.com)
  • Immunoprecipitation assays detect the interaction of a target protein with other proteins or nucleic acids. (genscript.com)
  • Chromatin I mmunoprecipitation (ChIP) is a type of immunoprecipitation used to investigate regions of genome associated with a target DNA-binding protein, or conversely to identify specific proteins associated with a particular region of the genome. (genscript.com)
  • The chromatin immunoprecipitation (ChIP) method provides an ideal tool for detecting direct or indirect interactions between proteins of interest and DNAs with known sequences. (sciencemag.org)
  • To probe the rewiring of signaling pathways and the heterogeneity across individual cancer cells, we developed a single-cell version of the co-immunoprecipitation (co-IP) analysis that examines the amount and PPIs of target proteins immunoprecipitated from individual cells. (rsc.org)
  • There are many methods, such as yeast two-hybrid, co-immunoprecipitation and so on, for determining the interaction between two proteins. (thefreedictionary.com)
  • Chromatin Immunoprecipitation (ChIP) is a powerful technique for mapping the in vivo distribution of proteins associated with chromosomal DNA. (emdmillipore.com)
  • We applied a novel strategy to identify immunogenic proteins of C. pneumoniae TW183 combining metabolic radiolabeling of de novo-synthesized chlamydial antigens with immunoprecipitation. (asm.org)
  • To verify the performance and specificity of Thermo Scientific antibodies, we have created a comprehensive workflow to assess antibody specificity using immunoprecipitation combined with mass spectrometry (IP-MS). In preliminary experiments, we screened more than 500 antibodies to nearly 100 key cancer signaling proteins expressed across 12 cultured tumor cell lines. (labroots.com)
  • Cross-linking Chromatin Immunoprecipitation (ChIP) has become a popular method to detect the in vivo binding of proteins to DNA. (epigenome-noe.net)
  • Chromatin Immunoprecipitation (ChIP) is an important technique allowing the researcher to analyze in vivo interactions of proteins with genomic DNA. (merckmillipore.com)
  • Assay for total protein then adjust concentration to approximately 1 mg/ml with Immunoprecipitation Buffer. (neb.com)
  • Assay reproducibility was initially assessed by random selection of the ANA 3+ or 4+ sera number of serum specimens tested and repeating the immunoprecipitation assay. (cdc.gov)
  • These seemingly complex issues can be directly addressed by a powerful methodology termed the chromatin immunoprecipitation (ChIP) assay. (nih.gov)
  • OBJECTIVE To compare a recently developed immunoprecipitation assay (IPA) to the mouse protection bioassay (MPB), currently considered the "gold standard", for detecting antibodies against botulinum toxin A (BTX-A) and to correlate these assay results with clinical responses to BTX-A injections. (bmj.com)
  • 18 The primary aim of this study was to compare the MPB with a more recent immunoprecipitation assay (IPA) developed by Palace et al 19 and to correlate the presence of antibodies detected by these two assays to the patients' clinical response to BTX-A injections. (bmj.com)
  • After modification of a chromatin immunoprecipitation assay, we cloned nine chromatin fragments which represent both strong and weak in vivo E2F binding sites. (asm.org)
  • Using an indirect immunofluorescence assay (IFA)-based approach and applying different immunoprecipitation (IP), chromatographic and mass spectrometric protocols was possible to isolate and identify a spectrum of autoantigens from brain tissue. (frontiersin.org)
  • DULIP: A Dual Luminescence-Based Co-Immunoprecipitation Assay for Interactome Mapping in Mammalian Cells. (thebiogrid.org)
  • Here, we present DULIP, a dual luminescence-based co-immunoprecipitation assay, for systematic PPI mapping in mammalian cells. (thebiogrid.org)
  • Porvair Sciences has published an informative 8-page brochure providing scientists with a background to Chromatin Immunoprecipitation (ChIP), an introduction to Chromatrap® ChIP assay technology and how Chromatrap® compares to traditional bead based methodologies. (technologynetworks.com)
  • Measurements of autoantibodies to interferon-ω (IFN-ω) in patients with autoimmune polyglandular syndrome type 1 (APS-1) were performed using a new immunoprecipitation assay (IPA) based on 125I-labeled IFN-ω. (ovid.com)
  • Sigma's Imprint brand of antibodies are validated in ChIP application and lot tested each time for successful chromatin immunoprecipitation experiment. (sigmaaldrich.com)
  • Alternatively, Protein G Magnetic Beads ( NEB #S1430S ) can be used for immunoprecipitations with monoclonal antibodies. (neb.com)
  • At BioLegend, we offer a wide-array of Go-ChIP-Grade™ antibodies and an enzymatic kit featuring a solid-state immunoprecipitation platform ideal for consistency between experiments. (biolegend.com)
  • After shearing the DNA into small fragments, specific DNA-protein complexes are isolated by immunoprecipitation with protein-specific ANTIBODIES. (umassmed.edu)
  • Co-immunoprecipitation was carried out in MCF-7/ADR whole cell lysates with protein G-Sepharose beads and antibodies against Anxa2 and P-gp. (nih.gov)
  • It is also suitable for indirect immunofluorescence (10-20μg/mL using HA-tagged fusion protein transfected cells), immunoprecipitation (2.5-4μg/test using HA-tagged fusion protein from cell lysates), and western blot (0.5-0.8μg/mL using HA-tagged fusion protein transfected cell extracts). (sigmaaldrich.com)
  • Testing of IgG autoantibodies to human cellular antigens was performed on samples that showed a 3+ or greater immunofluorescence staining using immunoprecipitation assays by a standard method (Reeves WH,et al. (cdc.gov)
  • a key transcriptional regulator), and mot-2 (a p53 inhibitor) and their relationships in arsenite-induced transformed HELF cells by two-dimensional electrophoresis, reverse-transcriptase polymerase chain reaction, Western blot, immunofluorescence, and co-immunoprecipitation assays. (thefreedictionary.com)
  • The interaction between P-gp and Anxa2 was examined by immunoprecipitation and immunofluorescence confocal microscopy analyses. (nih.gov)
  • Automation of IP protocol -learn how the Thermo Scientific KingFisher platform now supports immunoprecipitation automation. (thermofisher.com)
  • The Imprint ChIP Kit provides a complete solution for Chromatin Immunoprecipitation including columns and reagents for DNA purification and an integrated protocol for ChIP DNA amplification with our GenomePlex ® Whole Genome Amplification Kit ( WGA2 ). (sigmaaldrich.com)
  • Comparison of time required for protocol completion from fixation through purification using different Chromatin Immunoprecipitation kits. (sigmaaldrich.com)
  • Use 25 µl of Protein A or Protein G Magnetic Beads per 200 µl of crude cell lysate containing 200-500 µg of total protein in a standard immunoprecipitation protocol. (neb.com)
  • Serum samples were tested using the standard immunoprecipitation protocol (Reeves WH,et al. (cdc.gov)
  • Read our ChIP protocol and watch our video protocol to learn more about lysate preparation, chromatin cross-linking, DNA shearing by sonication, immunoprecipitation, reverse cross-linking, DNA purification, DNA amplification, and DNA quantification for ChIP. (novusbio.com)
  • This protocol offers a general guideline for immunoprecipitation with GenScript MagBeads . (genscript.com)
  • A detailed protocol on how to perform chromatin immunoprecipitation in solid tissue samples. (utexas.edu)
  • 2006) Based on these studies we set up a dual cross-linking ChIP protocol that we have successfully employed to improve immunoprecipitation of complexes in which tested factors are not in direct contact with DNA. (epigenome-noe.net)
  • Here we provide a robust Chromatin Immunoprecipitation (ChIP) protocol, allowing in vivo analyses of multiple chromatin modifications and binding of histone modifiers in different plant organs and tissues. (uzh.ch)
  • In this paper, we report a simplified and detailed protocol for robust immunoprecipitation of A beta in brain tissue prior to mass spectrometric detection exemplified by a study using transgenic mice. (diva-portal.org)
  • G-Biosciences Cross-Linking Immunoprecipitation kit contains all the reagents necessary to complete all aspects of immunoprecipitation, with the exception of labeling. (gbiosciences.com)
  • Single day chromatin immunoprecipitation (ChIP) kit containing all necessary reagents to enable ChIP from low input amounts of chromatin obtained from either cells or tissues using magnetic A/G beads. (emdmillipore.com)
  • Single day chromatin immunoprecipitation (ChIP) kit containing all necessary reagents to perform 22 individual chromatin immunoprecipitation (ChIP) reactions using magnetic A beads. (merckmillipore.com)
  • Major immunoprecipitation vendors include Thermo Fisher Scientific (US), Abcam (UK), Bio-Rad Laboratories (US), Merck KGaA (Germany), GenScript. (marketsandmarkets.com)
  • This up-to-date volume includes protocols that illustrate the broad use of chromatin immunoprecipitation (ChIP) and ChIP-related methods in a variety of biological research areas. (springer.com)
  • Authoritative and practical, Chromatin Immunoprecipitation: Methods and Protocols features techniques, including bioinformatic analysis of ChIP data, will be of interest to a very broad research community in the fields of biochemistry, molecular biology, microbiology, and biomedicine. (springer.com)
  • We invite you to browse all of our products approved for Immunoprecipitation (IP), ChIP and similar methods. (antibodies-online.com)
  • Immunoaffinity chromatographic and immunoprecipitation methods combined with mass spectrometry for characterization of circulating transthyretin. (diva-portal.org)
  • We have developed methods which enable us to identify en masse , in vivo targets of RBPs such as HuR from cell lines and now for the first time, solid tissues as well. (aacrjournals.org)
  • Two related methods that allow in vivo identification of transcription factor binding sites are chromatin immunoprecipitation (ChIP) and chromatin affinity purification (ChAP). For ChAP, the protein of interest is tagged with a peptide or protein, which can be used for affinity purification of the protein-DNA complex and hence, the identification of the target gene. (biomedcentral.com)
  • At this point the immunoprecipitation is performed resulting in the purification of protein-DNA complexes. (wikipedia.org)
  • The flexible format allows for immunoprecipitation and purification of DNA from mammalian cells or tissue in a convenient strip-well format. (sigmaaldrich.com)
  • immunoprecipitation and micro-scale antibody purification. (genscript.com)
  • Analysis of Histone Modifications in Rodent Pancreatic Islets by Native Chromatin Immunoprecipitation. (nih.gov)
  • This makes it possible for users to perform Chromatin immunoprecipitation (ChIP) assays using only 1000 cells per immunoprecipitation. (news-medical.net)
  • Traditional immunoprecipitation (IP) uses agarose beads coated in protein A/G. Most of these IP protocols require more than three washing steps to eliminate background and non-specific binding. (genscript.com)
  • ChIP-seq, or chromatin immunoprecipitation and sequencing, is a technique that allows researchers to understand transcriptional regulation via mapping of protein-DNA interactions and epigenetic markers on a genome-wide scale. (news-medical.net)
  • We developed a method, ChIP-sequencing (ChIP-seq), combining chromatin immunoprecipitation (ChIP) and massively parallel sequencing to identify mammalian DNA sequences bound by transcription factors in vivo . (nature.com)
  • The Ion Proton System combined with the MAGnify Chromatin Immunoprecipitation System delivers fast, reliable, and high-quality ChIP-Seq results at an affordable price. (thermofisher.com)
  • Following lysis of cross-linked cells and immunoprecipitation of bacterial RNA polymerase, DNA associated with enriched RNA polymerase was hybridized to probes corresponding to different regions of known genes to determine the in vivo distribution and density of RNA polymerase at these genes. (wikipedia.org)
  • Excess detergent in the lysis buffer may inhibit antibody binding during immunoprecipitation, so lysis buffer volume and composition must be optimized. (biolegend.com)
  • The SCW Buffer is unique to the Magna ChIP HiSens kit and enables the use of a single buffer for multiple steps of the ChIP process (sonication, chromatin immunoprecipitation, and wash). (emdmillipore.com)
  • Publication trends show that the fastest growing method for immunoprecipitation incorporates the use of magnetic beads and that Invitrogen Dynabeads products are the most used and referenced products in this trend. (thermofisher.com)
  • All magnetic beads immunoprecipitation wholesalers & magnetic beads immunoprecipitation manufacturers come from members. (lightneasy.org)
  • We doesn't provide magnetic beads immunoprecipitation products or service, please contact them directly and verify their companies info carefully. (lightneasy.org)
  • Immunoprecipitation and wash steps are fast and easy with magnetic beads because the pellet forms on the side of the tube, even pulling completely out of the buffer. (activemotif.com)
  • The use of magnetic beads has recently gained popularity as a quicker, more accurate approach for immunoprecipitation. (genscript.com)
  • Immobilized [3H]acetylated N-acetyltransferase-His8Flag intermediate cannot be detected in immunoprecipitation experiments with anti-Flag M2 beads. (figshare.com)
  • Immunoprecipitation ( IP ) is the technique of precipitating a protein antigen out of solution using an antibody that specifically binds to that particular protein. (wikipedia.org)
  • The combined procedures of immunoprecipitation and SDS-PAGE can be a powerful tool to assess the amount and size of an antibody-reactive antigen present in a complex protein mixture. (rockland-inc.com)
  • It is known that immunoprecipitation is a method to test target protein or antigen in solution. (biology-online.org)
  • Every method to immunoprecipitation has its certain operation process, but most of them are based on the specificity between antibody and antigen. (biology-online.org)
  • Hands-on activity of immunoprecipitation and isolate specific antigen and antibody. (astralscientific.com.au)
  • Sequential immunoprecipitation performed at different stages of the chlamydial developmental cycle revealed that the 60- to 62-kDa antigen is strongly upregulated after 24 to 48 h of host cell infection and is presented as a major immunogen in both C. pneumoniae -infected patients and mice. (asm.org)
  • Tiling arrays are increasingly used in chromatin immunoprecipitation (IP) experiments (ChIP on chip). (harvard.edu)
  • Chromatin immunoprecipitation (ChIP) is a powerful research technique used to identify and analyze protein-DNA interactions within the genome in vivo . (cellsignal.com)
  • The RNA immunoprecipitation (RIP) technique has been used to identify a large number of Hfq-associated RNA species, and can be effectively used to detect Hfq-bound RNAs in vivo, which can be subsequently analyzed with Northern blotting, quantitative PCR, microarray analysis, or deep sequencing. (springer.com)
  • Measurement of protein-DNA interactions in vivo by chromatin immunoprecipitation. (nih.gov)
  • Chromatin Immunoprecipitation determines the in vivo chromatin binding sites of a transcription factor or other protein of interest. (openwetware.org)
  • Site-specific chromatin immunoprecipitation: a selective method to individually analyze neighboring transcription factor binding sites in vivo. (biomedsearch.com)
  • A widely used method to analyze TFBSs in vivo is the chromatin immunoprecipitation (ChIP). (biomedsearch.com)
  • This alteration enables the specific immunoprecipitation and individual examination of occupied sites, even in a complex system of adjacent binding motifs in vivo. (biomedsearch.com)
  • The in vivo association of histone H1 with specific genes in Tetrahymena thermophila was studied by using a simplified cross-linking and immunoprecipitation technique. (asm.org)
  • This technique, in which no cross linking is used, allows one to en masse identify different in vivo mRNA targets which different RBPs interact with. (aacrjournals.org)
  • ChIP-IT Express Kits include a strong bar magnet that can be used to turn pipette tip boxes into magnetic stands for immunoprecipitation and washing. (activemotif.com)
  • Remnant liver tissue was homogenized and analyzed by co-immunoprecipitation and western blotting. (thefreedictionary.com)
  • In short, the method involves the fixation of plant tissue and the isolation of the total protein-DNA mixture, followed by an immunoprecipitation step with an antibody directed against the protein of interest. (biomedcentral.com)
  • This kit allows for quick and reproducible immunoprecipitation (IP) by using a 96 well filter plate. (bio-medicine.org)
  • Take 1000 ug whole cell lysate, add 2 ug of immunoprecipitation antibody. (avivasysbio.com)
  • Here, endogenously formed Hfq-RNA complexes of Y. pestis were captured with immunoprecipitation with a monoclonal anti-Flag antibody. (springer.com)
  • Slurries were prepared for immunoprecipitation using 100 µl chromatin stock from each extraction, depending on the number of cells: 1000, 10,000, 100,000. (news-medical.net)
  • 2) Lyse the cells with 0.5 ml cold Immunoprecipitation Buffer (150 mM NaCl, 10 mM Tris-HCl (pH 7.4), 1 mM EDTA, 1 mM EGTA (pH 8.0), 0.2 mM sodium ortho-vanadate, 0.2 mM PMSF, 1% Triton X-100, 0.5% NP-40). (neb.com)
  • 3.5 million cells are required for each immunoprecipitation (IP) (IgG control and specific antibody). (bio-protocol.org)
  • Co-immunoprecipitation of Anxa2 and P-gp in MCF-7/ADR cells. (nih.gov)
  • A high-throughput chromatin immunoprecipitation approach reveals principles of dynamic gene regulation in mammals. (broadinstitute.org)
  • Here, we develop a high-throughput Chromatin ImmunoPrecipitation (HT-ChIP) method to systematically map protein-DNA interactions. (broadinstitute.org)
  • The chromatin is sonicated to generate small fragments, and an immunoprecipitation is conducted with an antibody against the desired factor or histone modification. (nih.gov)
  • After the DNA is sheared, the DNA fragments that interact with the protein of interest are isolated by immunoprecipitation. (creativebiomart.net)
  • Fujita, T. and Fujii, H. (2012) Efficient isolation of specific genomic regions by insertional chromatin immunoprecipitation (iChIP) with a second-generation tagged LexA DNA-binding domain. (scirp.org)
  • Isolation of protein-DNA interactions by chromatin immunoprecipitation (ChIP) followed by massively parallel DNA sequencing is an unbiased method for identifying and quantitating DNA sites that are associated to a chromatin-bound protein(s) of interest in any cell type or organism with known genomic sequence. (thermofisher.com)
  • Chromatin immunoprecipitation (ChIP) is a method used to determine the location of DNA binding sites on the genome for a particular protein of interest. (wikipedia.org)
  • Co-Immunoprecipitation (Co-IP) is a powerful method that is most widely used by researchers to analyze protein-protein interactions. (genscript.com)
  • Hoshino, A. and Fujii, H. (2009) Insertional chromatin immunoprecipitation: a method for isolating specific genomic regions. (scirp.org)
  • Chromatin immunoprecipitation (ChIP) has become an important and popular method for the study of DNA-protein interactions. (creativebiomart.net)
  • We conclude that, due to its high sensitivity and concurrent preservation of conformational epitopes, metabolic radiolabeling of chlamydial antigens combined with immunoprecipitation may be a useful method to reveal important immunogens in respiratory C. pneumoniae infection which might have been missed by immunoblot analysis. (asm.org)
  • Appropriate isotype controls are highly recommended in order to show specific binding in your primary antibody immunoprecipitation. (cellsignal.com)
  • Fujita, T. and Fujii, H. (2011) Direct idenification of insulator components by insertional chromatin immunoprecipitation. (scirp.org)
  • Immunoprecipitation requires that the antibody be coupled to a solid substrate at some point in the procedure. (wikipedia.org)
  • Using the anti-HTR12 antibody we developed a chromatin immunoprecipitation (ChIP) procedure to determine which centromeric repeats, if any, are incorporated into the centromere/kinetochore complex in Arabidopsis. (genetics.org)
  • This chapter aims to describe one such technique, that of co-immunoprecipitation. (springer.com)
  • Join our presenters as they discuss how to identify protein interactions though immunoprecipitation and overcome biochemical problems that can be associated with this technique. (abcam.com)
  • Co-immunoprecipitation (co-IP) has become a standard technique, but its protein-band output provides only static, qualitative information about protein-protein interactions. (nih.gov)
  • We developed a chromatin immunoprecipitation (ChIP) technique using an antibody against the centromeric H3 histone, HTR12, in Arabidopsis. (genetics.org)
  • Immunoprecipitation is a routinely used technique that removes a protein or peptide, which specifically reacts with an antibody, from a solution. (astralscientific.com.au)
  • Immunoprecipitation Technique For 6 groups of 4-5 or 24-30 students. (astralscientific.com.au)
  • Successful chromatin immunoprecipitation requires an effective, specific, and high quality antibody. (sigmaaldrich.com)
  • RNA Immunoprecipitation (RIP) is performed with an antibody that targets a specific RNA-binding protein. (genscript.com)
  • ChIP methodology often involves protein-DNA and protein-protein cross-linking, fragmentation of the cross-linked chromatin, and subsequent immunoprecipitation of chromatin with an antibody specific to a target protein. (emdmillipore.com)
  • Additional targets of CodY, a GTP-activated repressor of early stationary-phase genes in Bacillus subtilis , were identified by combining chromatin immunoprecipitation, DNA microarray hybridization, and gel mobility shift assays. (asm.org)
  • Formaldehyde cross-linking and immunoprecipitation demonstrate developmental changes in H1 association with transcriptionally active genes. (asm.org)
  • Immunoprecipitation and subsequent isolation of nucleic acids allows for the investigation of protein:nucleic acid interactions. (bio-protocol.org)
  • Has anybody had any experience (good or bad) using the advertised Clontech polyclonal GFP antibody for immunoprecipitations or Western blots? (bio.net)