Antibodies: 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).Antibody Specificity: 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.Antibodies, Viral: Immunoglobulins produced in response to VIRAL ANTIGENS.Antibodies, Monoclonal: Antibodies produced by a single clone of cells.Antibody Formation: The production of ANTIBODIES by proliferating and differentiated B-LYMPHOCYTES under stimulation by ANTIGENS.Antibodies, Neutralizing: Antibodies that reduce or abolish some biological activity of a soluble antigen or infectious agent, usually a virus.Antibody Affinity: A measure of the binding strength between antibody and a simple hapten or antigen determinant. It depends on the closeness of stereochemical fit between antibody combining sites and antigen determinants, on the size of the area of contact between them, and on the distribution of charged and hydrophobic groups. It includes the concept of "avidity," which refers to the strength of the antigen-antibody bond after formation of reversible complexes.Fluorescent Antibody Technique: 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.Antibodies, Anti-Idiotypic: Antibodies which react with the individual structural determinants (idiotopes) on the variable region of other antibodies.Binding Sites, Antibody: Local surface sites on antibodies which react with antigen determinant sites on antigens (EPITOPES.) They are formed from parts of the variable regions of FAB FRAGMENTS.Phosphoric Monoester Hydrolases: A group of hydrolases which catalyze the hydrolysis of monophosphoric esters with the production of one mole of orthophosphate. EC 3.1.3.HIV Antibodies: Antibodies reactive with HIV ANTIGENS.Enamel Organ: Epithelial cells surrounding the dental papilla and differentiated into three layers: the inner enamel epithelium, consisting of ameloblasts which eventually form the enamel, and the enamel pulp and external enamel epithelium, both of which atrophy and disappear before and upon eruption of the tooth, respectively.Epitopes: Sites on an antigen that interact with specific antibodies.Antibodies, Neoplasm: Immunoglobulins induced by antigens specific for tumors other than the normally occurring HISTOCOMPATIBILITY ANTIGENS.Antibodies, Protozoan: Immunoglobulins produced in a response to PROTOZOAN ANTIGENS.Antibodies, Antinuclear: Autoantibodies directed against various nuclear antigens including DNA, RNA, histones, acidic nuclear proteins, or complexes of these molecular elements. Antinuclear antibodies are found in systemic autoimmune diseases including systemic lupus erythematosus, Sjogren's syndrome, scleroderma, polymyositis, and mixed connective tissue disease.Phosphorylation: The introduction of a phosphoryl group into a compound through the formation of an ester bond between the compound and a phosphorus moiety.Molecular Sequence Data: 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.Cross Reactions: Serological reactions in which an antiserum against one antigen reacts with a non-identical but closely related antigen.Amino Acid Sequence: 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.Tooth Calcification: The process whereby calcium salts are deposited in the dental enamel. The process is normal in the development of bones and teeth. (Boucher's Clinical Dental Terminology, 4th ed, p43)Hypophosphatasia: A genetic metabolic disorder resulting from serum and bone alkaline phosphatase deficiency leading to hypercalcemia, ethanolamine phosphatemia, and ethanolamine phosphaturia. Clinical manifestations include severe skeletal defects resembling vitamin D-resistant rickets, failure of the calvarium to calcify, dyspnea, cyanosis, vomiting, constipation, renal calcinosis, failure to thrive, disorders of movement, beading of the costochondral junction, and rachitic bone changes. (From Dorland, 27th ed)Dentinogenesis: The formation of dentin. Dentin first appears in the layer between the ameloblasts and odontoblasts and becomes calcified immediately. Formation progresses from the tip of the papilla over its slope to form a calcified cap becoming thicker by the apposition of new layers pulpward. A layer of uncalcified dentin intervenes between the calcified tissue and the odontoblast and its processes. (From Jablonski, Dictionary of Dentistry, 1992)Immunoglobulin M: A class of immunoglobulin bearing mu chains (IMMUNOGLOBULIN MU-CHAINS). IgM can fix COMPLEMENT. The name comes from its high molecular weight and originally being called a macroglobulin.Autoantibodies: Antibodies that react with self-antigens (AUTOANTIGENS) of the organism that produced them.Antibodies, Fungal: Immunoglobulins produced in a response to FUNGAL ANTIGENS.Neutralization Tests: The measurement of infection-blocking titer of ANTISERA by testing a series of dilutions for a given virus-antiserum interaction end-point, which is generally the dilution at which tissue cultures inoculated with the serum-virus mixtures demonstrate cytopathology (CPE) or the dilution at which 50% of test animals injected with serum-virus mixtures show infectivity (ID50) or die (LD50).Oxygen Isotopes: Stable oxygen atoms that have the same atomic number as the element oxygen, but differ in atomic weight. O-17 and 18 are stable oxygen isotopes.Alkaline Phosphatase: An enzyme that catalyzes the conversion of an orthophosphoric monoester and water to an alcohol and orthophosphate. EC 3.1.3.1.Antigen-Antibody Reactions: The processes triggered by interactions of ANTIBODIES with their ANTIGENS.Antibodies, Bispecific: Antibodies, often monoclonal, in which the two antigen-binding sites are specific for separate ANTIGENIC DETERMINANTS. They are artificial antibodies produced by chemical crosslinking, fusion of HYBRIDOMA cells, or by molecular genetic techniques. They function as the main mediators of targeted cellular cytotoxicity and have been shown to be efficient in the targeting of drugs, toxins, radiolabeled haptens, and effector cells to diseased tissue, primarily tumors.Single-Chain Antibodies: A form of antibodies consisting only of the variable regions of the heavy and light chains (FV FRAGMENTS), connected by a small linker peptide. They are less immunogenic than complete immunoglobulin and thus have potential therapeutic use.Apatites: A group of phosphate minerals that includes ten mineral species and has the general formula X5(YO4)3Z, where X is usually calcium or lead, Y is phosphorus or arsenic, and Z is chlorine, fluorine, or OH-. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)Mice, Inbred BALB CAntibodies, Blocking: Antibodies that inhibit the reaction between ANTIGEN and other antibodies or sensitized T-LYMPHOCYTES (e.g., antibodies of the IMMUNOGLOBULIN G class that compete with IGE antibodies for antigen, thereby blocking an allergic response). Blocking antibodies that bind tumors and prevent destruction of tumor cells by CYTOTOXIC T-LYMPHOCYTES have also been called enhancing antibodies. (Rosen et al., Dictionary of Immunology, 1989)Immunoglobulin G: The major immunoglobulin isotype class in normal human serum. There are several isotype subclasses of IgG, for example, IgG1, IgG2A, and IgG2B.Tooth Germ: The collective tissues from which an entire tooth is formed, including the DENTAL SAC; ENAMEL ORGAN; and DENTAL PAPILLA. (From Jablonski, Dictionary of Dentistry, 1992)Antigen-Antibody Complex: 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.Kinetics: The rate dynamics in chemical or physical systems.Blotting, Western: 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.Immunoglobulin Fab Fragments: Univalent antigen-binding fragments composed of one entire IMMUNOGLOBULIN LIGHT CHAIN and the amino terminal end of one of the IMMUNOGLOBULIN HEAVY CHAINS from the hinge region, linked to each other by disulfide bonds. Fab contains the IMMUNOGLOBULIN VARIABLE REGIONS, which are part of the antigen-binding site, and the first IMMUNOGLOBULIN CONSTANT REGIONS. This fragment can be obtained by digestion of immunoglobulins with the proteolytic enzyme PAPAIN.Antibodies, Heterophile: Antibodies elicited in a different species from which the antigen originated. These antibodies are directed against a wide variety of interspecies-specific antigens, the best known of which are Forssman, Hanganutziu-Deicher (H-D), and Paul-Bunnell (P-B). Incidence of antibodies to these antigens--i.e., the phenomenon of heterophile antibody response--is useful in the serodiagnosis, pathogenesis, and prognosis of infection and latent infectious states as well as in cancer classification.Antibodies, Catalytic: Antibodies that can catalyze a wide variety of chemical reactions. They are characterized by high substrate specificity and share many mechanistic features with enzymes.Cell Line: Established cell cultures that have the potential to propagate indefinitely.Lansoprazole: A 2,2,2-trifluoroethoxypyridyl derivative of timoprazole that is used in the therapy of STOMACH ULCERS and ZOLLINGER-ELLISON SYNDROME. The drug inhibits H(+)-K(+)-EXCHANGING ATPASE which is found in GASTRIC PARIETAL CELLS. Lansoprazole is a racemic mixture of (R)- and (S)-isomers.Recombinant Proteins: Proteins prepared by recombinant DNA technology.Immunohistochemistry: Histochemical localization of immunoreactive substances using labeled antibodies as reagents.Cells, Cultured: 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.Rabbits: 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.Immunoglobulin A: Represents 15-20% of the human serum immunoglobulins, mostly as the 4-chain polymer in humans or dimer in other mammals. Secretory IgA (IMMUNOGLOBULIN A, SECRETORY) is the main immunoglobulin in secretions.Phosphofructokinase-2: An allosteric enzyme that regulates glycolysis and gluconeogenesis by catalyzing the transfer of a phosphate group from ATP to fructose-6-phosphate to yield fructose-2,6-bisphosphate, an allosteric effector for the other 6-phosphofructokinase, PHOSPHOFRUCTOKINASE-1. Phosphofructokinase-2 is bifunctional: the dephosphorylated form is a kinase and the phosphorylated form is a phosphatase that breaks down fructose-2,6-bisphosphate to yield fructose-6-phosphate.Antibodies, Monoclonal, Humanized: Antibodies from non-human species whose protein sequences have been modified to make them nearly identical with human antibodies. If the constant region and part of the variable region are replaced, they are called humanized. If only the constant region is modified they are called chimeric. INN names for humanized antibodies end in -zumab.Fluorescent Antibody Technique, Indirect: A form of fluorescent antibody technique commonly used to detect serum antibodies and immune complexes in tissues and microorganisms in specimens from patients with infectious diseases. The technique involves formation of an antigen-antibody complex which is labeled with fluorescein-conjugated anti-immunoglobulin antibody. (From Bennington, Saunders Dictionary & Encyclopedia of Laboratory Medicine and Technology, 1984)Diphosphates: Inorganic salts of phosphoric acid that contain two phosphate groups.Molecular Weight: The sum of the weight of all the atoms in a molecule.Hybridomas: Cells artificially created by fusion of activated lymphocytes with neoplastic cells. The resulting hybrid cells are cloned and produce pure MONOCLONAL ANTIBODIES or T-cell products, identical to those produced by the immunologically competent parent cell.Immune Sera: 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.PhosphoproteinsEpitope Mapping: Methods used for studying the interactions of antibodies with specific regions of protein antigens. Important applications of epitope mapping are found within the area of immunochemistry.Antibodies, Antiphospholipid: Autoantibodies directed against phospholipids. These antibodies are characteristically found in patients with systemic lupus erythematosus (LUPUS ERYTHEMATOSUS, SYSTEMIC;), ANTIPHOSPHOLIPID SYNDROME; related autoimmune diseases, some non-autoimmune diseases, and also in healthy individuals.Immunization: Deliberate stimulation of the host's immune response. ACTIVE IMMUNIZATION involves administration of ANTIGENS or IMMUNOLOGIC ADJUVANTS. PASSIVE IMMUNIZATION involves administration of IMMUNE SERA or LYMPHOCYTES or their extracts (e.g., transfer factor, immune RNA) or transplantation of immunocompetent cell producing tissue (thymus or bone marrow).Ameloblasts: Cylindrical epithelial cells in the innermost layer of the ENAMEL ORGAN. Their functions include contribution to the development of the dentinoenamel junction by the deposition of a layer of the matrix, thus producing the foundation for the prisms (the structural units of the DENTAL ENAMEL), and production of the matrix for the enamel prisms and interprismatic substance. (From Jablonski's Dictionary of Dentistry, 1992)Alveolar Process: The thickest and spongiest part of the maxilla and mandible hollowed out into deep cavities for the teeth.Signal Transduction: 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.Substrate Specificity: A characteristic feature of enzyme activity in relation to the kind of substrate on which the enzyme or catalytic molecule reacts.Immunoenzyme Techniques: Immunologic techniques based on the use of: (1) enzyme-antibody conjugates; (2) enzyme-antigen conjugates; (3) antienzyme antibody followed by its homologous enzyme; or (4) enzyme-antienzyme complexes. These are used histologically for visualizing or labeling tissue specimens.Diphosphoglyceric AcidsPhosphotransferases: A rather large group of enzymes comprising not only those transferring phosphate but also diphosphate, nucleotidyl residues, and others. These have also been subdivided according to the acceptor group. (From Enzyme Nomenclature, 1992) EC 2.7.Antigens, Bacterial: Substances elaborated by bacteria that have antigenic activity.Enzyme-Linked Immunosorbent Assay: 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.Antigens, Surface: 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.2-Pyridinylmethylsulfinylbenzimidazoles: Compounds that contain benzimidazole joined to a 2-methylpyridine via a sulfoxide linkage. Several of the compounds in this class are ANTI-ULCER AGENTS that act by inhibiting the POTASSIUM HYDROGEN ATPASE found in the PROTON PUMP of GASTRIC PARIETAL CELLS.Binding Sites: The parts of a macromolecule that directly participate in its specific combination with another molecule.Immunization, Passive: Transfer of immunity from immunized to non-immune host by administration of serum antibodies, or transplantation of lymphocytes (ADOPTIVE TRANSFER).Protein Binding: 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.Protein Conformation: The characteristic 3-dimensional shape of a protein, including the secondary, supersecondary (motifs), tertiary (domains) and quaternary structure of the peptide chain. PROTEIN STRUCTURE, QUATERNARY describes the conformation assumed by multimeric proteins (aggregates of more than one polypeptide chain).Models, Molecular: Models used experimentally or theoretically to study molecular shape, electronic properties, or interactions; includes analogous molecules, computer-generated graphics, and mechanical structures.Immunoassay: A technique using antibodies for identifying or quantifying a substance. Usually the substance being studied serves as antigen both in antibody production and in measurement of antibody by the test substance.Immunoglobulin Fragments: Partial immunoglobulin molecules resulting from selective cleavage by proteolytic enzymes or generated through PROTEIN ENGINEERING techniques.Mice, Inbred C57BLAntigens, Viral: Substances elaborated by viruses that have antigenic activity.Immunoblotting: 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.Radioimmunoassay: Classic quantitative assay for detection of antigen-antibody reactions using a radioactively labeled substance (radioligand) either directly or indirectly to measure the binding of the unlabeled substance to a specific antibody or other receptor system. Non-immunogenic substances (e.g., haptens) can be measured if coupled to larger carrier proteins (e.g., bovine gamma-globulin or human serum albumin) capable of inducing antibody formation.Electrophoresis, Polyacrylamide Gel: Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.B-Lymphocytes: Lymphoid cells concerned with humoral immunity. They are short-lived cells resembling bursa-derived lymphocytes of birds in their production of immunoglobulin upon appropriate stimulation.Complement Fixation Tests: Serologic tests based on inactivation of complement by the antigen-antibody complex (stage 1). Binding of free complement can be visualized by addition of a second antigen-antibody system such as red cells and appropriate red cell antibody (hemolysin) requiring complement for its completion (stage 2). Failure of the red cells to lyse indicates that a specific antigen-antibody reaction has taken place in stage 1. If red cells lyse, free complement is present indicating no antigen-antibody reaction occurred in stage 1.Hemagglutination Tests: Sensitive tests to measure certain antigens, antibodies, or viruses, using their ability to agglutinate certain erythrocytes. (From Stedman, 26th ed)Hemagglutination Inhibition Tests: Serologic tests in which a known quantity of antigen is added to the serum prior to the addition of a red cell suspension. Reaction result is expressed as the smallest amount of antigen which causes complete inhibition of hemagglutination.Sequence Alignment: The arrangement of two or more amino acid or base sequences from an organism or organisms in such a way as to align areas of the sequences sharing common properties. The degree of relatedness or homology between the sequences is predicted computationally or statistically based on weights assigned to the elements aligned between the sequences. This in turn can serve as a potential indicator of the genetic relatedness between the organisms.Antibodies, Antineutrophil Cytoplasmic: Autoantibodies directed against cytoplasmic constituents of POLYMORPHONUCLEAR LEUKOCYTES and/or MONOCYTES. They are used as specific markers for GRANULOMATOSIS WITH POLYANGIITIS and other diseases, though their pathophysiological role is not clear. ANCA are routinely detected by indirect immunofluorescence with three different patterns: c-ANCA (cytoplasmic), p-ANCA (perinuclear), and atypical ANCA.Immunoglobulin Variable Region: That region of the immunoglobulin molecule that varies in its amino acid sequence and composition, and comprises the binding site for a specific antigen. It is located at the N-terminus of the Fab fragment of the immunoglobulin. It includes hypervariable regions (COMPLEMENTARITY DETERMINING REGIONS) and framework regions.Seroepidemiologic Studies: EPIDEMIOLOGIC STUDIES based on the detection through serological testing of characteristic change in the serum level of specific ANTIBODIES. Latent subclinical infections and carrier states can thus be detected in addition to clinically overt cases.Immunoglobulin Idiotypes: Unique genetically-controlled determinants present on ANTIBODIES whose specificity is limited to a single group of proteins (e.g., another antibody molecule or an individual myeloma protein). The idiotype appears to represent the antigenicity of the antigen-binding site of the antibody and to be genetically codetermined with it. The idiotypic determinants have been precisely located to the IMMUNOGLOBULIN VARIABLE REGION of both immunoglobin polypeptide chains.Pyrophosphatases: A group of enzymes within the class EC 3.6.1.- that catalyze the hydrolysis of diphosphate bonds, chiefly in nucleoside di- and triphosphates. They may liberate either a mono- or diphosphate. EC 3.6.1.-.T-Lymphocytes: Lymphocytes responsible for cell-mediated immunity. Two types have been identified - cytotoxic (T-LYMPHOCYTES, CYTOTOXIC) and helper T-lymphocytes (T-LYMPHOCYTES, HELPER-INDUCER). They are formed when lymphocytes circulate through the THYMUS GLAND and differentiate to thymocytes. When exposed to an antigen, they divide rapidly and produce large numbers of new T cells sensitized to that antigen.Immunologic Techniques: Techniques used to demonstrate or measure an immune response, and to identify or measure antigens using antibodies.Serine: A non-essential amino acid occurring in natural form as the L-isomer. It is synthesized from GLYCINE or THREONINE. It is involved in the biosynthesis of PURINES; PYRIMIDINES; and other amino acids.Protein Kinases: A family of enzymes that catalyze the conversion of ATP and a protein to ADP and a phosphoprotein.Antigens, Neoplasm: Proteins, glycoprotein, or lipoprotein moieties on surfaces of tumor cells that are usually identified by monoclonal antibodies. Many of these are of either embryonic or viral origin.Base Sequence: The sequence of PURINES and PYRIMIDINES in nucleic acids and polynucleotides. It is also called nucleotide sequence.Vascular Calcification: Deposition of calcium into the blood vessel structures. Excessive calcification of the vessels are associated with ATHEROSCLEROTIC PLAQUES formation particularly after MYOCARDIAL INFARCTION (see MONCKEBERG MEDIAL CALCIFIC SCLEROSIS) and chronic kidney diseases which in turn increase VASCULAR STIFFNESS.Immunosorbent Techniques: Techniques for removal by adsorption and subsequent elution of a specific antibody or antigen using an immunosorbent containing the homologous antigen or antibody.Haptens: Small antigenic determinants capable of eliciting an immune response only when coupled to a carrier. Haptens bind to antibodies but by themselves cannot elicit an antibody response.Threonine: An essential amino acid occurring naturally in the L-form, which is the active form. It is found in eggs, milk, gelatin, and other proteins.Databases, Protein: Databases containing information about PROTEINS such as AMINO ACID SEQUENCE; PROTEIN CONFORMATION; and other properties.Hardness: The mechanical property of material that determines its resistance to force. HARDNESS TESTS measure this property.Antibody Diversity: The phenomenon of immense variability characteristic of ANTIBODIES. It enables the IMMUNE SYSTEM to react specifically against the essentially unlimited kinds of ANTIGENS it encounters. Antibody diversity is accounted for by three main theories: (1) the Germ Line Theory, which holds that each antibody-producing cell has genes coding for all possible antibody specificities, but expresses only the one stimulated by antigen; (2) the Somatic Mutation Theory, which holds that antibody-producing cells contain only a few genes, which produce antibody diversity by mutation; and (3) the Gene Rearrangement Theory, which holds that antibody diversity is generated by the rearrangement of IMMUNOGLOBULIN VARIABLE REGION gene segments during the differentiation of the ANTIBODY-PRODUCING CELLS.Cattle: 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.Peptide Library: A collection of cloned peptides, or chemically synthesized peptides, frequently consisting of all possible combinations of amino acids making up an n-amino acid peptide.RNA, Messenger: 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.Hepatitis C Antibodies: Antibodies to the HEPATITIS C ANTIGENS including antibodies to envelope, core, and non-structural proteins.Isoantibodies: Antibodies from an individual that react with ISOANTIGENS of another individual of the same species.Immunoglobulin Isotypes: The classes of immunoglobulins found in any species of animal. In man there are nine classes that migrate in five different groups in electrophoresis; they each consist of two light and two heavy protein chains, and each group has distinguishing structural and functional properties.Fractures, Spontaneous: Fractures occurring as a result of disease of a bone or from some undiscoverable cause, and not due to trauma. (Dorland, 27th ed)Flow Cytometry: Technique using an instrument system for making, processing, and displaying one or more measurements on individual cells obtained from a cell suspension. Cells are usually stained with one or more fluorescent dyes specific to cell components of interest, e.g., DNA, and fluorescence of each cell is measured as it rapidly transverses the excitation beam (laser or mercury arc lamp). Fluorescence provides a quantitative measure of various biochemical and biophysical properties of the cell, as well as a basis for cell sorting. Other measurable optical parameters include light absorption and light scattering, the latter being applicable to the measurement of cell size, shape, density, granularity, and stain uptake.Immunoglobulins: Multi-subunit proteins which function in IMMUNITY. They are produced by B LYMPHOCYTES from the IMMUNOGLOBULIN GENES. They are comprised of two heavy (IMMUNOGLOBULIN HEAVY CHAINS) and two light chains (IMMUNOGLOBULIN LIGHT CHAINS) with additional ancillary polypeptide chains depending on their isoforms. The variety of isoforms include monomeric or polymeric forms, and transmembrane forms (B-CELL ANTIGEN RECEPTORS) or secreted forms (ANTIBODIES). They are divided by the amino acid sequence of their heavy chains into five classes (IMMUNOGLOBULIN A; IMMUNOGLOBULIN D; IMMUNOGLOBULIN E; IMMUNOGLOBULIN G; IMMUNOGLOBULIN M) and various subclasses.Phosphoric Diester Hydrolases: A class of enzymes that catalyze the hydrolysis of one of the two ester bonds in a phosphodiester compound. EC 3.1.4.Organogenesis: Formation of differentiated cells and complicated tissue organization to provide specialized functions.Antibodies, Monoclonal, Murine-Derived: Antibodies obtained from a single clone of cells grown in mice or rats.Glycoproteins: Conjugated protein-carbohydrate compounds including mucins, mucoid, and amyloid glycoproteins.Vaccination: Administration of vaccines to stimulate the host's immune response. This includes any preparation intended for active immunological prophylaxis.Time Factors: Elements of limited time intervals, contributing to particular results or situations.Hepatitis B Antibodies: Antibodies to the HEPATITIS B ANTIGENS, including antibodies to the surface (Australia) and core of the Dane particle and those to the "e" antigens.Immunodiffusion: Technique involving the diffusion of antigen or antibody through a semisolid medium, usually agar or agarose gel, with the result being a precipitin reaction.Sensitivity and Specificity: Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed)Binding, Competitive: The interaction of two or more substrates or ligands with the same binding site. The displacement of one by the other is used in quantitative and selective affinity measurements.Sequence Homology, Amino Acid: The degree of similarity between sequences of amino acids. This information is useful for the analyzing genetic relatedness of proteins and species.Peptide Fragments: Partial proteins formed by partial hydrolysis of complete proteins or generated through PROTEIN ENGINEERING techniques.X-Ray Microtomography: X-RAY COMPUTERIZED TOMOGRAPHY with resolution in the micrometer range.Immunity, Maternally-Acquired: Resistance to a disease-causing agent induced by the introduction of maternal immunity into the fetus by transplacental transfer or into the neonate through colostrum and milk.Insulin Antibodies: Antibodies specific to INSULIN.Complement System Proteins: Serum glycoproteins participating in the host defense mechanism of COMPLEMENT ACTIVATION that creates the COMPLEMENT MEMBRANE ATTACK COMPLEX. Included are glycoproteins in the various pathways of complement activation (CLASSICAL COMPLEMENT PATHWAY; ALTERNATIVE COMPLEMENT PATHWAY; and LECTIN COMPLEMENT PATHWAY).Lupus Erythematosus, Systemic: A chronic, relapsing, inflammatory, and often febrile multisystemic disorder of connective tissue, characterized principally by involvement of the skin, joints, kidneys, and serosal membranes. It is of unknown etiology, but is thought to represent a failure of the regulatory mechanisms of the autoimmune system. The disease is marked by a wide range of system dysfunctions, an elevated erythrocyte sedimentation rate, and the formation of LE cells in the blood or bone marrow.Incisor: Any of the eight frontal teeth (four maxillary and four mandibular) having a sharp incisal edge for cutting food and a single root, which occurs in man both as a deciduous and a permanent tooth. (Jablonski, Dictionary of Dentistry, 1992, p820)Spleen: An encapsulated lymphatic organ through which venous blood filters.Enzyme Inhibitors: Compounds or agents that combine with an enzyme in such a manner as to prevent the normal substrate-enzyme combination and the catalytic reaction.Autoantigens: Endogenous tissue constituents that have the ability to interact with AUTOANTIBODIES and cause an immune response.Recombinant Fusion Proteins: 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.Tyrosine: A non-essential amino acid. In animals it is synthesized from PHENYLALANINE. It is also the precursor of EPINEPHRINE; THYROID HORMONES; and melanin.Precipitin Tests: 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.Species Specificity: The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.Molar: The most posterior teeth on either side of the jaw, totaling eight in the deciduous dentition (2 on each side, upper and lower), and usually 12 in the permanent dentition (three on each side, upper and lower). They are grinding teeth, having large crowns and broad chewing surfaces. (Jablonski, Dictionary of Dentistry, 1992, p821)Antigens, Protozoan: Any part or derivative of any protozoan that elicits immunity; malaria (Plasmodium) and trypanosome antigens are presently the most frequently encountered.Peptides: 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.Serologic Tests: Diagnostic procedures involving immunoglobulin reactions.Antibody-Dependent Cell Cytotoxicity: The phenomenon of antibody-mediated target cell destruction by non-sensitized effector cells. The identity of the target cell varies, but it must possess surface IMMUNOGLOBULIN G whose Fc portion is intact. The effector cell is a "killer" cell possessing Fc receptors. It may be a lymphocyte lacking conventional B- or T-cell markers, or a monocyte, macrophage, or polynuclear leukocyte, depending on the identity of the target cell. The reaction is complement-independent.Apoptosis: 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.Magnetic Resonance Spectroscopy: Spectroscopic method of measuring the magnetic moment of elementary particles such as atomic nuclei, protons or electrons. It is employed in clinical applications such as NMR Tomography (MAGNETIC RESONANCE IMAGING).Single-Domain Antibodies: An immunoglobulin fragment composed of one variable domain from an IMMUNOGLOBULIN HEAVY CHAIN or IMMUNOGLOBULIN LIGHT CHAIN.Polysaccharides, Bacterial: Polysaccharides found in bacteria and in capsules thereof.Chromatography, Affinity: 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)Iodine Radioisotopes: Unstable isotopes of iodine that decay or disintegrate emitting radiation. I atoms with atomic weights 117-139, except I 127, are radioactive iodine isotopes.Adenosine Triphosphate: An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.Bacterial Vaccines: Suspensions of attenuated or killed bacteria administered for the prevention or treatment of infectious bacterial disease.Cloning, Molecular: 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.Mice, Inbred Strains: Genetically identical individuals developed from brother and sister matings which have been carried out for twenty or more generations, or by parent x offspring matings carried out with certain restrictions. All animals within an inbred strain trace back to a common ancestor in the twentieth generation.Immunochemistry: 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.Viral Envelope Proteins: Layers of protein which surround the capsid in animal viruses with tubular nucleocapsids. The envelope consists of an inner layer of lipids and virus specified proteins also called membrane or matrix proteins. The outer layer consists of one or more types of morphological subunits called peplomers which project from the viral envelope; this layer always consists of glycoproteins.Immunoglobulin Heavy Chains: The largest of polypeptide chains comprising immunoglobulins. They contain 450 to 600 amino acid residues per chain, and have molecular weights of 51-72 kDa.Tissue Distribution: Accumulation of a drug or chemical substance in various organs (including those not relevant to its pharmacologic or therapeutic action). This distribution depends on the blood flow or perfusion rate of the organ, the ability of the drug to penetrate organ membranes, tissue specificity, protein binding. The distribution is usually expressed as tissue to plasma ratios.Proto-Oncogene Proteins c-akt: A protein-serine-threonine kinase that is activated by PHOSPHORYLATION in response to GROWTH FACTORS or INSULIN. It plays a major role in cell metabolism, growth, and survival as a core component of SIGNAL TRANSDUCTION. Three isoforms have been described in mammalian cells.Dose-Response Relationship, Immunologic: A specific immune response elicited by a specific dose of an immunologically active substance or cell in an organism, tissue, or cell.Proteomics: The systematic study of the complete complement of proteins (PROTEOME) of organisms.Antigens, CD: Differentiation antigens residing on mammalian leukocytes. CD stands for cluster of differentiation, which refers to groups of monoclonal antibodies that show similar reactivity with certain subpopulations of antigens of a particular lineage or differentiation stage. The subpopulations of antigens are also known by the same CD designation.Tumor Cells, Cultured: 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.Membrane Glycoproteins: Glycoproteins found on the membrane or surface of cells.Membrane Proteins: 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.Extracellular Matrix: A meshwork-like substance found within the extracellular space and in association with the basement membrane of the cell surface. It promotes cellular proliferation and provides a supporting structure to which cells or cell lysates in culture dishes adhere.Radioimmunotherapy: Radiotherapy where cytotoxic radionuclides are linked to antibodies in order to deliver toxins directly to tumor targets. Therapy with targeted radiation rather than antibody-targeted toxins (IMMUNOTOXINS) has the advantage that adjacent tumor cells, which lack the appropriate antigenic determinants, can be destroyed by radiation cross-fire. Radioimmunotherapy is sometimes called targeted radiotherapy, but this latter term can also refer to radionuclides linked to non-immune molecules (see RADIOTHERAPY).Lymphocyte Activation: Morphologic alteration of small B LYMPHOCYTES or T LYMPHOCYTES in culture into large blast-like cells able to synthesize DNA and RNA and to divide mitotically. It is induced by INTERLEUKINS; MITOGENS such as PHYTOHEMAGGLUTININS, and by specific ANTIGENS. It may also occur in vivo as in GRAFT REJECTION.Erythrocytes: Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.Viral Vaccines: Suspensions of attenuated or killed viruses administered for the prevention or treatment of infectious viral disease.Lymphocytes: White blood cells formed in the body's lymphoid tissue. The nucleus is round or ovoid with coarse, irregularly clumped chromatin while the cytoplasm is typically pale blue with azurophilic (if any) granules. Most lymphocytes can be classified as either T or B (with subpopulations of each), or NATURAL KILLER CELLS.Immunoelectrophoresis: A technique that combines protein electrophoresis and double immunodiffusion. In this procedure proteins are first separated by gel electrophoresis (usually agarose), then made visible by immunodiffusion of specific antibodies. A distinct elliptical precipitin arc results for each protein detectable by the antisera.Microscopy, Electron: Microscopy using an electron beam, instead of light, to visualize the sample, thereby allowing much greater magnification. The interactions of ELECTRONS with specimens are used to provide information about the fine structure of that specimen. In TRANSMISSION ELECTRON MICROSCOPY the reactions of the electrons that are transmitted through the specimen are imaged. In SCANNING ELECTRON MICROSCOPY an electron beam falls at a non-normal angle on the specimen and the image is derived from the reactions occurring above the plane of the specimen.Immunoglobulin E: An immunoglobulin associated with MAST CELLS. Overexpression has been associated with allergic hypersensitivity (HYPERSENSITIVITY, IMMEDIATE).Epitopes, B-Lymphocyte: Antigenic determinants recognized and bound by the B-cell receptor. Epitopes recognized by the B-cell receptor are located on the surface of the antigen.Cell Culture Techniques: Methods for maintaining or growing CELLS in vitro.Immunoglobulin Light Chains: Polypeptide chains, consisting of 211 to 217 amino acid residues and having a molecular weight of approximately 22 kDa. There are two major types of light chains, kappa and lambda. Two Ig light chains and two Ig heavy chains (IMMUNOGLOBULIN HEAVY CHAINS) make one immunoglobulin molecule.Agglutination Tests: Tests that are dependent on the clumping of cells, microorganisms, or particles when mixed with specific antiserum. (From Stedman, 26th ed)Protein-Serine-Threonine Kinases: A group of enzymes that catalyzes the phosphorylation of serine or threonine residues in proteins, with ATP or other nucleotides as phosphate donors.Vaccines, Synthetic: Small synthetic peptides that mimic surface antigens of pathogens and are immunogenic, or vaccines manufactured with the aid of recombinant DNA techniques. The latter vaccines may also be whole viruses whose nucleic acids have been modified.Enzyme Activation: 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.Immunotherapy: Manipulation of the host's immune system in treatment of disease. It includes both active and passive immunization as well as immunosuppressive therapy to prevent graft rejection.Swine: Any of various animals that constitute the family Suidae and comprise stout-bodied, short-legged omnivorous mammals with thick skin, usually covered with coarse bristles, a rather long mobile snout, and small tail. Included are the genera Babyrousa, Phacochoerus (wart hogs), and Sus, the latter containing the domestic pig (see SUS SCROFA).Microscopy, Immunoelectron: Microscopy in which the samples are first stained immunocytochemically and then examined using an electron microscope. Immunoelectron microscopy is used extensively in diagnostic virology as part of very sensitive immunoassays.HIV-1: The type species of LENTIVIRUS and the etiologic agent of AIDS. It is characterized by its cytopathic effect and affinity for the T4-lymphocyte.Disease Models, Animal: Naturally occurring or experimentally induced animal diseases with pathological processes sufficiently similar to those of human diseases. They are used as study models for human diseases.Immunotoxins: Semisynthetic conjugates of various toxic molecules, including RADIOACTIVE ISOTOPES and bacterial or plant toxins, with specific immune substances such as IMMUNOGLOBULINS; MONOCLONAL ANTIBODIES; and ANTIGENS. The antitumor or antiviral immune substance carries the toxin to the tumor or infected cell where the toxin exerts its poisonous effect.Antiphospholipid Syndrome: The presence of antibodies directed against phospholipids (ANTIBODIES, ANTIPHOSPHOLIPID). The condition is associated with a variety of diseases, notably systemic lupus erythematosus and other connective tissue diseases, thrombopenia, and arterial or venous thromboses. In pregnancy it can cause abortion. Of the phospholipids, the cardiolipins show markedly elevated levels of anticardiolipin antibodies (ANTIBODIES, ANTICARDIOLIPIN). Present also are high levels of lupus anticoagulant (LUPUS COAGULATION INHIBITOR).Radioimmunodetection: Use of radiolabeled antibodies for diagnostic imaging of neoplasms. Antitumor antibodies are labeled with diverse radionuclides including iodine-131, iodine-123, indium-111, or technetium-99m and injected into the patient. Images are obtained by a scintillation camera.Chickens: Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA.Escherichia coli: A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.HIV Envelope Protein gp120: External envelope protein of the human immunodeficiency virus which is encoded by the HIV env gene. It has a molecular weight of 120 kDa and contains numerous glycosylation sites. Gp120 binds to cells expressing CD4 cell-surface antigens, most notably T4-lymphocytes and monocytes/macrophages. Gp120 has been shown to interfere with the normal function of CD4 and is at least partly responsible for the cytopathic effect of HIV.Cell Adhesion: Adherence of cells to surfaces or to other cells.beta 2-Glycoprotein I: A 44-kDa highly glycosylated plasma protein that binds phospholipids including CARDIOLIPIN; APOLIPOPROTEIN E RECEPTOR; membrane phospholipids, and other anionic phospholipid-containing moieties. It plays a role in coagulation and apoptotic processes. Formerly known as apolipoprotein H, it is an autoantigen in patients with ANTIPHOSPHOLIPID ANTIBODIES.Hindlimb: Either of two extremities of four-footed non-primate land animals. It usually consists of a FEMUR; TIBIA; and FIBULA; tarsals; METATARSALS; and TOES. (From Storer et al., General Zoology, 6th ed, p73)DNA: 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).Immunoglobulin A, Secretory: The principle immunoglobulin in exocrine secretions such as milk, respiratory and intestinal mucin, saliva and tears. The complete molecule (around 400 kD) is composed of two four-chain units of IMMUNOGLOBULIN A, one SECRETORY COMPONENT and one J chain (IMMUNOGLOBULIN J-CHAINS).Hemocyanin

Rapid and sensitive identification of epitope-containing peptides by direct matrix-assisted laser desorption/ionization tandem mass spectrometry of peptides affinity-bound to antibody beads. (1/86)

A method has been developed for rapid and sensitive identification of epitope-containing peptides, based on direct MALDI-MS/MS analysis of epitope-containing peptides affinity bound to affinity beads. This technique provides sequence information of the epitope that allows unambiguous identification of the epitope either by database searching or de novo sequencing. With MALDI-MS, affinity beads with bound peptides can be placed directly on the MALDI target and analyzed. Coupling a MALDI source to an orthogonal injection quadrupole time-of-flight (QqTOF) mass spectrometer allows direct sequencing of the bound peptides. In contrast to ESI-MS/MS, elution of the affinity-bound peptides followed by additional concentration and purification steps is not required, thus reducing the potential for sample loss. Direct mass spectrometric sequencing of affinity-bound peptides eliminates the need for chemical or enzymatic sequencing. Other advantages of this direct MALDI-MS/MS analysis of epitope-containing peptides bound to the affinity beads include its sensitivity (femtomole levels) and speed. In addition, direct analysis of peptides on affinity beads does not adversely affect the high mass accuracy of a QqTOF, and database searching can be performed on the MS/MS spectra obtained. In proof-of-principle experiments, this method has been demonstrated on beads containing immobilized antibodies against phosphotyrosine, the c-myc epitope tag, as well as immobilized avidin. Furthermore, de novo sequencing of epitope-containing peptides is demonstrated. The first application of this method was with anti-FLAG-tag affinity beads, where direct MALDI MS/MS was used to determine an unexpected enzymatic cleavage site on a growth factor protein.  (+info)

Phosphorylation state-specific antibodies: applications in investigative and diagnostic pathology. (2/86)

Until recently, the investigation of protein phosphorylation was limited to biochemical studies of enzyme activities in homogenized tissues. The availability of hundreds of phosphorylation state-specific antibodies (PSSAs) now makes possible the study of protein phosphorylation in situ, and is opening many exciting opportunities in investigative and diagnostic pathology. This review illustrates the power of PSSAs, especially in immunohistochemical applications to human disease and animal models. Technical considerations, including antibody specificity and lability of phosphoepitopes, are covered, along with potential pitfalls, illustrated by a case study. In the arena of oncology, PSSAs may prove especially valuable in directly demonstrating the efficacy of chemotherapies targeted at protein kinase cascades. Novel applications of PSSAs are also beginning to reveal molecular mechanisms of inflammatory, degenerative, and toxin-induced diseases.  (+info)

Differential phosphorylation and subcellular localization of La RNPs associated with precursor tRNAs and translation-related mRNAs. (3/86)

The La protein facilitates the production of tRNAs in the nucleus and the translation of specific mRNAs in the cytoplasm. We report that human La that is phosphorylated on serine 366 (pLa) is nucleoplasmic and associated with precursor tRNAs and other nascent RNA polymerase III transcripts while nonphosphorylated (np)La is cytoplasmic and associated with a subset of mRNAs that contain 5'-terminal oligopyrimidine (5'TOP) motifs known to control protein synthesis. Thus, La ribonucleoproteins (RNP) exist in distinct states that differ in subcellular localization, serine 366 phosphorylation, and associated RNAs. These results are consistent with a model in which the relative concentrations of the La S366 isoforms in different subcellular compartments in conjunction with the relative concentrations of specific RNA ligands in these compartments determine the differential association of npLa and pLa with their respective classes of associated RNAs.  (+info)

Replication protein A (RPA) phosphorylation prevents RPA association with replication centers. (4/86)

Mammalian replication protein A (RPA) undergoes DNA damage-dependent phosphorylation at numerous sites on the N terminus of the RPA2 subunit. To understand the functional significance of RPA phosphorylation, we expressed RPA2 variants in which the phosphorylation sites were converted to aspartate (RPA2(D)) or alanine (RPA2(A)). Although RPA2(D) was incorporated into RPA heterotrimers and supported simian virus 40 DNA replication in vitro, the RPA2(D) mutant was selectively unable to associate with replication centers in vivo. In cells containing greatly reduced levels of endogenous RPA2, RPA2(D) again did not localize to replication sites, indicating that the defect in supporting chromosomal DNA replication is not due to competition with the wild-type protein. Use of phosphospecific antibodies demonstrated that endogenous hyperphosphorylated RPA behaves similarly to RPA2(D). In contrast, under DNA damage or replication stress conditions, RPA2(D), like RPA2(A) and wild-type RPA2, was competent to associate with DNA damage foci as determined by colocalization with gamma-H2AX. We conclude that RPA2 phosphorylation prevents RPA association with replication centers in vivo and potentially serves as a marker for sites of DNA damage.  (+info)

Biochemical characterization of the Drosophila wingless signaling pathway based on RNA interference. (5/86)

Regulation of Armadillo (Arm) protein levels through ubiquitin-mediated degradation plays a central role in the Wingless (Wg) signaling. Although zeste-white3 (Zw3)-mediated Arm phosphorylation has been implicated in its degradation, we have recently shown that casein kinase Ialpha (CKIalpha) also phosphorylates Arm and induces its degradation. However, it remains unclear how CKIalpha and Zw3, as well as other components of the Arm degradation complex, regulate Arm phosphorylation in response to Wg. In particular, whether Wg signaling suppresses CKIalpha- or Zw3-mediated Arm phosphorylation in vivo is unknown. To clarify these issues, we performed a series of RNA interference (RNAi)-based analyses in Drosophila S2R+ cells by using antibodies that specifically recognize Arm phosphorylated at different serine residues. These analyses revealed that Arm phosphorylation at serine-56 and at threonine-52, serine-48, and serine-44, is mediated by CKIalpha and Zw3, respectively, and that Zw3-directed Arm phosphorylation requires CKIalpha-mediated priming phosphorylation. Daxin stimulates Zw3- but not CKIalpha-mediated Arm phosphorylation. Wg suppresses Zw3- but not CKIalpha-mediated Arm phosphorylation, indicating that a vital regulatory step in Wg signaling is Zw3-mediated Arm phosphorylation. In addition, further RNAi-based analyses of the other aspects of the Wg pathway clarified that Wg-induced Dishevelled phosphorylation is due to CKIalpha and that presenilin and protein kinase A play little part in the regulation of Arm protein levels in Drosophila tissue culture cells.  (+info)

Site-selective regulation of platelet-derived growth factor beta receptor tyrosine phosphorylation by T-cell protein tyrosine phosphatase. (6/86)

The platelet-derived growth factor (PDGF) beta receptor mediates mitogenic and chemotactic signals. Like other tyrosine kinase receptors, the PDGF beta receptor is negatively regulated by protein tyrosine phosphatases (PTPs). To explore whether T-cell PTP (TC-PTP) negatively regulates the PDGF beta receptor, we compared PDGF beta receptor tyrosine phosphorylation in wild-type and TC-PTP knockout (ko) mouse embryos. PDGF beta receptors were hyperphosphorylated in TC-PTP ko embryos. Fivefold-higher ligand-induced receptor phosphorylation was observed in TC-PTP ko mouse embryo fibroblasts (MEFs) as well. Reexpression of TC-PTP partly abolished this difference. As determined with site-specific phosphotyrosine antibodies, the extent of hyperphosphorylation varied among different autophosphorylation sites. The phospholipase Cgamma1 binding site Y1021, previously implicated in chemotaxis, displayed the largest increase in phosphorylation. The increase in Y1021 phosphorylation was accompanied by increased phospholipase Cgamma1 activity and migratory hyperresponsiveness to PDGF. PDGF beta receptor tyrosine phosphorylation in PTP-1B ko MEFs but not in PTPepsilon ko MEFs was also higher than that in control cells. This increase occurred with a site distribution different from that seen after TC-PTP depletion. PDGF-induced migration was not increased in PTP-1B ko cells. In summary, our findings identify TC-PTP as a previously unrecognized negative regulator of PDGF beta receptor signaling and support the general notion that PTPs display site selectivity in their action on tyrosine kinase receptors.  (+info)

ZIP kinase is responsible for the phosphorylation of myosin II and necessary for cell motility in mammalian fibroblasts. (7/86)

Reorganization of actomyosin is an essential process for cell migration and myosin regulatory light chain (MLC20) phosphorylation plays a key role in this process. Here, we found that zipper-interacting protein (ZIP) kinase plays a predominant role in myosin II phosphorylation in mammalian fibroblasts. Using two phosphorylation site-specific antibodies, we demonstrated that a significant portion of the phosphorylated MLC20 is diphosphorylated and that the localization of mono- and diphosphorylated myosin is different from each other. The kinase responsible for the phosphorylation was ZIP kinase because (a) the kinase in the cell extracts phosphorylated Ser19 and Thr18 of MLC20 with similar potency; (b) immunodepletion of ZIP kinase from the cell extracts markedly diminished its myosin II kinase activity; and (c) disruption of ZIP kinase expression by RNA interference diminished myosin phosphorylation, and resulted in the defect of cell polarity and migration efficiency. These results suggest that ZIP kinase is critical for myosin phosphorylation and necessary for cell motile processes in mammalian fibroblasts.  (+info)

Molecular pharmacology and antitumor activity of PX-866, a novel inhibitor of phosphoinositide-3-kinase signaling. (8/86)

We have developed biologically stable semisynthetic viridins as inhibitors of phosphoinositide (PtdIns)-3-kinases. The most active compound was PX-866 (acetic acid (1S,4E,10R,11R,13S,14R)-[4-diallylaminomethylene-6-hydroxy-1-methoxymethyl-10,13- dimethyl-3,7,17-trioxo-1,3,4,7,10,11,12,13,14,15,16,17-dodecahydro-2-oxa-cyclopen ta[a]phenanthren-11-yl ester), which inhibited purified PtdIns-3-kinase with an IC50 of 0.1 nmol/L and PtdIns-3-kinase signaling measured by phospho-Ser473-Akt levels in HT-29 colon cancer cells with an IC50 of 20 nmol/L. PX-866 administered to mice at 10 mg/kg inhibited phospho-Ser473-Akt in HT-29 colon tumor xenografts up to 80% with recovery taking >48 hours after p.o. administration but more rapidly after i.v. or i.p. administration. PX-866 was eliminated from mouse plasma with a half-life of 18 minutes and a clearance of 360 mL/min/kg following i.v. administration and, when administered i.p. or p.o., showed first-pass metabolism with sequential N-deallylation. Synthetic standards of the N-deallylated metabolites of PX-866 inhibited PtdIns-3-kinase at low nanomolar per liter concentrations. PX-866 exhibited in vivo antitumor activity against s.c. OvCar-3 human ovarian cancer and A-549 human lung cancer xenografts in immunodeficient mice with log cell kills up to 1.2. PX-866 also increased the antitumor activity of cisplatin against A-549 xenografts and radiation treatment against OvCar-3 xenografts. The results show that PX-866 is a biologically stable broad-spectrum PtdIns-3-kinase inhibitor with good pharmacokinetics that causes prolonged inhibition of PtdIns-3-kinase signaling in human tumor xenografts. PX-866 exhibits single agent in vivo antitumor activity and increases the antitumor effects of cisplatin and radiation treatment.  (+info)

*Monoclonal antibody therapy

The advent of monoclonal antibody technology has made it possible to raise antibodies against specific antigens presented on ... However, Aβ concentration did not significantly change, along with other AD biomarkers, including phospho-tau expression, and ... The first FDA-approved therapeutic monoclonal antibody was a murine IgG2a CD3 specific transplant rejection drug, OKT3 (also ... Antibody-drug conjugates (ADCs) are antibodies linked to one or more drug molecules. Typically when the ADC meets the target ...

*Phosphoproteomics

Phosphopeptides are enriched using phosphospecific antibodies, immobilized metal affinity chromatography or titanium dioxide ( ... In one study design, cells are exposed to SILAC labelling and then stimulated by a specific growth factor. The cells are ... the current and simplest methods to enrich phosphoproteins are affinity purification using phosphospecific antibodies, ... Antiphosphotyrosine antibodies have been proven very successful in purification, but fewer reports have been published using ...

*DNA repair protein XRCC4

Anti-XRCC4 antibodies including phosphospecific antibodies to pS260 and pS318 in XRCC4 have been developed. Antibodies to XRCC4 ... As a result, this specific region of the genome is permanently lost and the deletion can lead to cancer and premature aging. ... "Anti-XRCC4 antibody - ChIP Grade (ab145) , Abcam". Abcam. ; "XRCC4 antibody , Western , SAB2102728". Sigma-Aldrich. Massip L, ... Antibodies are composed of two light and two heavy chains. The antigen binding site consists of two variable regions, VL and VH ...

*BRCA1

In vivo assessment using phospho-specific antibodies". J. Biol. Chem. 276 (20): 17276-80. doi:10.1074/jbc.M011681200. PMID ... There are a number of specific microRNAs, when overexpressed, that directly reduce expression of specific DNA repair proteins ( ... BRCA1 is a human tumor suppressor gene (to be specific, a caretaker gene), found in all humans; its protein, also called by the ... Yu X, Chini CC, He M, Mer G, Chen J (October 2003). "The BRCT domain is a phospho-protein binding domain". Science. 302 (5645 ...

*Ataxia telangiectasia and Rad3 related

In vivo assessment using phospho-specific antibodies". The Journal of Biological Chemistry. 276 (20): 17276-80. doi:10.1074/jbc ... ATR is a serine/threonine-specific protein kinase that is involved in sensing DNA damage and activating the DNA damage ... Furthermore, there are dramatic reductions with age in tissue-specific stem and progenitor cells, and exhaustion of tissue ...

*ATM serine/threonine kinase

In vivo assessment using phospho-specific antibodies". J. Biol. Chem. 276 (20): 17276-80. doi:10.1074/jbc.M011681200. PMID ... While deletion of the entire PRD domain abolishes the kinase activity of ATM, specific small deletions show no effect. Ataxia ...

*Phosphorylation

Such antibodies are called phospho-specific antibodies; hundreds of such antibodies are now available. They are becoming ... Antibodies can be used as powerful tool to detect whether a protein is phosphorylated at a particular site. Antibodies bind to ... The two enzymes have been identified as a specific glucokinase (ATP-D-glucose 6-phosphotransferase) and non-specific hexokinase ... In some very specific cases, the detection of the phosphorylation as a shift in the protein's electrophoretic mobility is ...

*EIF4EBP1

P sites detected by phospho-specific antibodies". J. Biol. Chem. 275 (43): 33836-33843. doi:10.1074/jbc.M006005200. PMID ... Pro motif and is activated by antibodies to a region near its COOH terminus". J. Biol. Chem. 272 (51): 32547-32550. doi:10.1074 ...

*Proteomics

... known as phospho-specific antibodies. Also, there are antibodies specific to other modifications. These can be used to ... Modified proteins can be studied by developing an antibody specific to that modification. For example, there are antibodies ... There are several specific techniques and protocols that use antibodies for protein detection. The enzyme-linked immunosorbent ... If a complex biological sample is analyzed, either a very specific antibody needs to be used in quantitative dot blot analysis ...

*Sigma-Aldrich

... phosphospecific antibodies, key signal transduction enzymes, and assay kits for cell signaling. ISOTEC provides isotopically ... SAGE also announced its first successful effort in creating a "knockout rabbit". Its facilities include a specific pathogen ...

*Reverse phase protein lysate microarray

Antibodies, especially phospho-specific reagents, often detect linear peptide sequences that may be masked due to the three- ... Strips with single band indicate specific antibodies that are suitable for RPMA use. Antibody performance should be also ... In addition, finding the appropriate antibody could require extensive screening of many antibodies by western blotting prior to ... both phospho-specific and total protein levels to be measured and analyzed at once. Once immunostaining has been performed ...

*UNC84B

... approach for identification of serine/threonine-phosphorylated proteins by enrichment with phospho-specific antibodies: ... Hartley JL, Temple GF, Brasch MA (2001). "DNA cloning using in vitro site-specific recombination". Genome Res. 10 (11): 1788-95 ...

*List of MeSH codes (D12.776)

... phospho-specific MeSH D12.776.377.715.548.114.252 - antibodies, protozoan MeSH D12.776.377.715.548.114.254 - antibodies, viral ... antibodies MeSH D12.776.377.715.548.114.071 - antibodies, anti-idiotypic MeSH D12.776.377.715.548.114.107 - antibodies, ... antibodies, bispecific MeSH D12.776.377.715.548.114.143 - antibodies, blocking MeSH D12.776.377.715.548.114.167 - antibodies, ... antibodies, helminth MeSH D12.776.377.715.548.114.191 - antibodies, heterophile MeSH D12.776.377.715.548.114.224 - antibodies, ...

*List of MeSH codes (D12.776.124)

... antibodies, neoplasm MeSH D12.776.124.486.485.114.248 -- antibodies, phospho-specific MeSH D12.776.124.486.485.114.252 -- ... antibodies, neoplasm MeSH D12.776.124.790.651.114.248 -- antibodies, phospho-specific MeSH D12.776.124.790.651.114.252 -- ... antibodies MeSH D12.776.124.486.485.114.071 -- antibodies, anti-idiotypic MeSH D12.776.124.486.485.114.089 -- antibodies, ... antibodies, blocking MeSH D12.776.124.486.485.114.167 -- antibodies, catalytic MeSH D12.776.124.486.485.114.179 -- antibodies, ...

*Signalway Antibody

... phospho-Ser37) ab and b-catenin(phospho-Tyr333) antibody. These antibodies have been shown to play an important role in cancer ... and modification-specific antibodies, as well as the Wnt/Notch/Hedgehog and AKT, Jak/Stat, MAPK, and TGFb/smads pathways. Yuhui ... Signalway Antibody is a biotechnology company based in College Park, Maryland, USA. It provides antibodies, proteins and kits ... Signalway Antibody LLC specializes in newly developed antibodies, including Bub3 Y207 ab, PKM2( ...

*Liposome

These targeting ligands could be monoclonal antibodies (making an immunoliposome), vitamins, or specific antigens, but must be ... They typically form after supplying enough energy to a dispersion of (phospho)lipids in a polar solvent, such as water, to ... Liposomes are presently being implemented for the specific oral delivery of certain dietary and nutritional supplements. A very ... "Specific targeting with poly (ethylene glycol)-modified liposomes: coupling of homing devices to the ends of the polymeric ...

*Chemical biology

By installing phospho-serine, phospho-threonine or analogous phosphonate mimics into native proteins, researchers are able to ... Functional metagenomic studies are designed to search for specific phenotypes that are associated with molecules with specific ... Both organic dyes and quantum dyes do not have the ability to recognize the protein of interest without the aid of antibodies, ... nonpolar) but not the specific identity of each amino acid in a sequence. Among other things, this strategy has been used to ...

*Caveolin 2

Phospho-caveolin-2 (Tyr(P)19) is localized near focal adhesions, remains associated with lipid rafts/caveolae, but no longer ... 1997). "Cell-type and tissue-specific expression of caveolin-2. Caveolins 1 and 2 co-localize and form a stable hetero- ... Characterization and epitope-mapping of a novel flotillin-1 monoclonal antibody probe". J. Biol. Chem. 274 (18): 12702-9. doi: ... Hartley JL, Temple GF, Brasch MA (2001). "DNA cloning using in vitro site-specific recombination". Genome Res. 10 (11): 1788-95 ...

*RELA

Specific modification patterns of RELA have also been observed in many cancer types. RELA may have a potential role as ... It is shown that ERα interacts with both p50 and RELA in vitro and in vivo, and RELA antibody can reduce ERα:ERE complex ... Phosphorylation at serine 276 in RELA allows its interaction with P-TEFb containing CDK9 and cyclin T1 subunits, and phospho- ... Cell-type-specific phosphorylation is also observed for RELA. Multiple-site phosphorylation is common in endothelial cells, and ...

*Polyomaviridae

"Association of Merkel cell polyomavirus-specific antibodies with Merkel cell carcinoma". Journal of the National Cancer ... The agnoprotein is a small multifunctional phospho-protein found in the late coding part of the genome of some polyomaviruses, ... Antibody assays are commonly used to detect presence of antibodies against individual viruses. Competition assays are ... Antibodies to the monkey lymphotropic polyomavirus have been detected in humans suggesting that this virus - or a closely ...

*Flow cytometry

Fluorophores, or simply "fluors", are typically attached to an antibody that recognizes a target feature on or in the cell; ... based upon the specific light scattering and fluorescent characteristics of each cell. It is a useful scientific instrument as ... phospho-proteins Scattering of light can be used to measure volume (by forward scatter) and morphological complexity (by side ... The amount of the analyte captured is detected via a biotinylated antibody against a secondary epitope of the protein, followed ...

*SISCAPA

... is captured by a sequence-specific anti-peptide antibody. The antibody, together with the captured target peptide, is then ... quantitative pharmacodynamic studies of phospho-signaling. 2015 May 18;:mcp.O115.050351. Anderson NL, Anderson NG, Pearson TW, ... A limitation of the specific peptide capture approach is the requirement for a specially-developed antibody against the ... Antibody-Coupled Magnetic Beads Can Be Reused in Immuno-MRM Assays To Reduce Cost and Extend Antibody Supply. J Proteome Res. ...

*Gel electrophoresis

A specific experiment example of an application of native gel electrophoresis is to check for enzymatic activity to verify the ... These can be transferred onto a nitrocellulose or PVDF membrane to be probed with antibodies and corresponding markers, such as ... it starts with the de-phosphorylation of 3-phospho-2-naphthoic acid-2'-4'-dimethyl aniline by alkaline phosphatase (water is ... In most cases, the gel is a crosslinked polymer whose composition and porosity is chosen based on the specific weight and ...

*Pick's disease

... but immunohistochemical staining using anti-tau and anti-ubiquitin antibodies have proven the most efficient and specific. ... CS1 maint: Uses authors parameter (link) Puig, B; Ferrer I; Ludueña RF; Avila J. (2005). "βII-tubulin and phospho-tau ... The second use of the term (and the one now used among professionals) is to mean a specific pathology that is one of the causes ... Numerous areas of the brain are affected by PiD, but the specific areas that are affected allow for differentiation between PiD ...

*EEF2K

Olsen JV, Blagoev B, Gnad F, Macek B, Kumar C, Mortensen P, Mann M (Nov 2006). "Global, in vivo, and site-specific ... autophosphorylation of Thr-348 leads to a conformational change in the kinase likely supported by the binding of phospho-Thr- ... antibodies in patients with systemic lupus erythematosus". Biochemical and Biophysical Research Communications. 293 (3): 1073-6 ...

*PIN1

Wulf G, Finn G, Suizu F, Lu KP (May 2005). "Phosphorylation-specific prolyl isomerization: is there an underlying theme?". ... isomerizes only phospho-Serine/Threonine-Proline motifs. The enzyme binds to a subset of proteins and thus plays a role as a ... "A hyperphosphorylated form of RNA polymerase II is the major interphase antigen of the phosphoprotein antibody MPM-2 and ... "The essential mitotic peptidyl-prolyl isomerase Pin1 binds and regulates mitosis-specific phosphoproteins". Genes & Development ...
Does anyone has experience with phospho-flow of peripheral blood samples? We would like to know how long after the blood draw the phosphorylation site remains stable. thanks in advance, kewal , Kewal Asosingh, Ph.D. , Department of Pathobiology/NC22 , Cleveland Clinic , 9500 Euclid Ave. , Cleveland, OH 44195 , , Phone: 216-444-0891 , Fax: 216-636-0104 =================================== P Please consider the environment before printing this e-mail Cleveland Clinic is ranked one of the top hospitals in America by U.S. News & World Report (2008). Visit us online at http://www.clevelandclinic.org for a complete listing of our services, staff and locations. Confidentiality Note: This message is intended for use only by the individual or entity to which it is addressed and may contain information that is privileged, confidential, and exempt from disclosure under applicable law. If the reader of this message is not the intended recipient or the employee or agent responsible for delivering the message ...
Phosphospecific peptide synthesis (up to 15 amino acids) -- 10 mg used for phosphospecific epitope affinity purification, 10 mg used for antibody production, and 10 mg delivered to client ...
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PPD® Laboratories biomarker lab has access to state-of-the-art instrumentation in both the bioanalytical lab and the central lab. The biomarker lab can accommodate research assays to fully validated assays with CAP/CLIA, GLP and GCLP compliance. The lab also provides training to clinical sites on proper sample processing for complex procedures, such as phospho-flow and activation assays. The biomarker lab has experience with various sample types including whole, frozen and lysed blood, as well as PBMCs. The test menu includes routine cell surface assays and intracellular assays.. Key Instrumentation: ...
T-Cell Receptor Signaling Phospho-Specific Array includes 188 highly specific and well-characterized phosphorylation antibodies in the T-cell receptor signaling pathway. (AA0040) - Products - Abnova
EGF Phospho-Specific Array includes 214 highly specific and well-characterized phosphorylation antibodies related to the EGF family of proteins. (AA0023) - Products - Abnova
SAB is a professional manufacturer of phospho-specific antibody products, committed to provide high-quality antibody research tools for the global life science researchers. Currently, SAB researches and develops more than 4000 kinds of antibody...
OUTLINE: Cryopreserved samples are incubated with cytokines (e.g., interleukins), growth factors (e.g., sargramostim [GM-CSF] and filgrastim [G-CSF]), chemotherapeutic agents (e.g., cytarabine, etoposide phosphate), and other modulators. Cells are fixed, permeabilized, and stained with antibodies that recognize extracellular markers in conjunction with intracellular activation-state specific epitopes of designated signaling molecules. Subsequently, cells are analyzed for multiparametric phospho-flow cytometry in a random manner (without knowledge of clinical variables and outcomes) to training and testing sets. Results are then compared with individual patient clinical outcomes. ...
It seemed that the state of tyrosine phosphorylation was not altered by the expression of the caveolin-1 mutant. We next used a variety of phospho-specific antibodies (12) that have been generated against the activated forms of well-known signal transducers. Fig. 4d ⇓ shows that both intracellular MAPKs and p38-MAPK 4 were constitutively activated in the caveolin-1 mutant clones. On the other hand, the status of phosphorylation of AKT was not changed, although v-Src-transformed cells (SR3Y1) showed increased phosphorylation of AKT. Again, these experiments demonstrated the capacity for the mutant to drive cell transformation. These observations are compatible with previous reports showing that alteration of caveolin-1 function could be involved in the cellular transformation as a dominant negative effect of caveolin-1.. The mutation-positive cases of breast cancer were mostly involved in the pathologically invasive types such as scirrhous carcinomas (18) . Hence, we suspected that the ...
The Janus kinase (JAK)/STAT signaling pathway is an active focus of MM research. In mononuclear cells from the BM of patients with MM, constitutively activated STAT3 signaling contributes to disease pathogenesis by preventing apoptosis [14]. MM cells express constitutively active forms of nuclear factor-κB and STAT3, and the suppression of these transcription factors inhibits the survival of these malignant cells [15]. Apoptosis has been shown to be induced in diverse MM cells treated with agents that inhibit the STAT3 signaling pathway [1617]. However, few studies have examined the clinical characteristics and prognosis of patients with BM tissue expression of PY-STAT3. Brown et al. [18] used phospho-flow cytometry to evaluate the constitutive expression of phosphorylated STAT3 (pSTAT3), pSTAT5, pERK, pAKT, and IL-6 receptor epitope in cryopreserved BM samples with respect to the clinical significance of positivity. In contrast to our results, the authors found no significant difference in OS ...
5-HETE stimulated PC3 cells to phosphorylate ERK1/2 and Akt, as detected in Western blots probed with phospho-specific antibodies (Fig. 2) ⇓ . Because blots probed with antibody to total (i.e., phosphorylated plus unphosphorylated species) ERK1/2 or Akt had no such change (data not shown), and because phosphorylation at the antibody-defined sites raises the activity of these kinases, Fig. 2 ⇓ data imply that 5-HETE induces ERK and Akt activation. ERK and Akt phosphorylation responses developed within 1 min, peaked at 5-10 min, and tended to persist for ≥60 min (ERK) or returned to baseline by 40 min (Akt). 5-HETE was more potent than 5-oxoETE, whereas 5-oxo-15-OH-ETE, 15(S)-hydroxy-6,8,11,13-Z,Z,Z,E-ETE, 8(S),12(S)-dihydroxy-5,9,11,13-Z,E,Z,E-ETE, and LTB4 were inactive (Fig. 2) ⇓ . 5-HETE, 5-oxoETE, and 5-oxo-15-OH-ETE likewise stimulated PMNs to activate ERKs; responses began by 1 min, peaked at 5-10 min, and declined thereafter. However, PMNs showed 5-fold rises in ERK phosphorylation ...
Figure 1 Antiphospho-Pak1 antibody specifically recognizes activated Pak1. (A) Activation loop sequence of the phosphorylated peptide used for production of polyclonal antiphospho-Pak1 antisera. (B) Immunoblots (lanes 1-4) and radiolabeled kinase reaction products (lanes 5 and 6) of baculovirus-produced Pak1. Purified KD or CA Pak1 was incubated in protein kinase buffer in the presence of ATP, then immunoblotted using anti-Pak1 (lanes 1 and 2) and antiphospho-Pak antisera (lanes 3 and 4) autoradiographed to detect antibody-bound proteins. Although both forms of the protein were detected with anti-Pak1, only activated Pak1 was detected using the phospho-specific antibody, despite the large amount of recombinant protein used in this assay. Lanes 5 and 6 show autoradiographed PAGE gel of reaction products from a kinase assay performed in the presence of [γ-32P]ATP. Molecular mass in kilodaltons is shown to the right of the blot. (C) Comparison between immunoblot and radiolabeled kinase reaction ...
Background: Heart failure (HF) is associated with decreased cardiac contractility and ventricular remodeling, features which are partially reversed by angiotensin converting enzyme inhibitors (ACEi). Since Rho kinase (ROCK) mediates cellular contraction through effects on the actin cytoskeleton and is an important regulator of cardiac fibrosis, we hypothesize that ACEi may improve heart function through inhibition of ROCK activity.. Methods and Results: We enrolled 30 consecutive HF subjects (average age: 52.7±8.2, 60% male in gender, NYHA class 3.2±1.1), with impaired left ventricular ejection fraction (LVEF) of less than 40% by echocardiography (average LVEF=32.1±8.2%), and age- and sex-matched 30 subjects with preserved LVEF function (average LVEF=65.4±9.4%) as the control group. ROCK activity was determined in peripheral leukocytes using a phospho-specific antibody for myosin binding subunit (MBS). Compared with the control group, the subjects with impaired LVEF have higher ROCK activity ...
A large body of work has implicated the activity of various kinases and phosphatases in the regulation of synaptic transmission by controlling the phosphorylation state of several synaptic proteins (for review, see Turner et al., 1999). To further understand the physiological significance of these modifications and to gain insights into their role in modulating synaptic strength and plasticity, we have generated a set of antibodies that specifically recognize synaptic proteins only in their phosphorylated form. In this study we report the biochemical characterization of the modulation of phosphorylation of rabphilin, a synaptic protein implicated in exocytosis, using phosphospecific antibodies directed against the two major phosphorylation sites of rabphilin, serine-234 and serine-274.. Our results show that the phosphorylation of rabphilin at serine-234 is greatly stimulated (about sevenfold over basal) by activation of PKA and high K+-induced membrane depolarization, a condition that mimics ...
Nucleases play important roles in DNA synthesis, recombination and repair. We have previously shown that human exonuclease 1 (hEXO1) is phosphorylated in response to agents stalling DNA replication and that hEXO1 consequently undergoes ubiquitination and degradation in a proteasome-dependent manner. In the present study, we have addressed the identity of the pathway transducing stalled-replication signals to hEXO1. Using chemical inhibitors, RNA interference, ATM- and ATR-deficient cell lines we have concluded that hEXO1 phosphorylation is ATR-dependent. By means of mass spectrometry, we have identified the sites of phosphorylation in hEXO1 in undamaged cells and in cells treated with hydroxyurea (HU). hEXO1 is phosphorylated at nine basal sites and three additional sites are induced by HU treatment. Analysis of single- and multiple-point mutants revealed that mutation to Ala of the three HU-induced sites of phosphorylation partially rescued HU-dependent degradation of hEXO1 and additionally ...
Histone deacetylase inhibitors (HDACi) have been reported to increase tumor antigen expression, and have been successfully tested as adjuvants for melanoma immunotherapy in mouse models. In this work, we tested the effects of a pan-HDACi on human lymphocytes and melanoma cell lines. Effects of the pan-HDACi panobinostat (LBH589) on cell viability, cell cycle, apoptosis and DNA damage were determined in peripheral blood mononuclear cells (PBMC) from two healthy donor (HD), thirteen patients with metastatic melanoma (MD), two bone marrow samples from different malignances, and twelve human melanoma cell lines. Intracellular signaling in lymphocytes, with or without cytokine stimulation, was analyzed by phospho-flow cytometry in one of each type. The 50% inhibition concentration (IC50) in PBMC was ,20 nM compared to ,600 nM in melanoma cell lines; ,40% apoptotic cell death in PBMC versus ,10% in melanoma cell lines was seen at the same concentration. Phospho-histone variant H2A.X (pH2A.X) increased ...
In article ,31D7ED66.3535 at ens-lyon.fr,, Stephane Ory ,Stephane.Ory at ens- lyon.fr, writes: , Does anyone know where antibody against phosphoserine and , phosphothreonine are available ? , , Thanks per advance Monoclonals are available from Sigma but when we tested them, they were not useful. Phosphoserine and phosphothreonine are about 100-1000X more abundant in proteins that phosphotyrosine so you need an additional level of specificity. A number of companies are selling antibodies to proteins that only detect those proteins if a particular ser/thr/tyr residue is phosphorylated. See NEB and UBI catalogues for example. Examples of phospho-specific antibodies to the following proteins: tau (microtubule-associated protein, various ser/thr) Jun (ser 63) STAT 3 (tyr 705) SEK1 ERK1/2 (tyr 204) p38 MAPK CREB (ser 133) Jim Woodgett Ontario Cancer Institute mailto:jwoodget at oci.utoronto.ca 610 University Avenue http://kinase.oci.utoronto.ca/woodgettlab.html Toronto, Ontario M5G 2M9 phone (416) ...
Bovine Serum Albumin (BSA) is used for various biochemical applications including ELISA (Enzyme-Linked Immunosorbent Assay), high content screening assays, western blotting, FACS Buffer and immunohistochemistry. BSA as a blocking reagent is particularly useful with casein-sensitive antibodies, such as phospho-specific antibodies. Also used as a nutrient in cell and microbial culture. In restriction digests, BSA is used to stabilize some enzymes during digestion of DNA and to prevent adhesion of the enzyme to reaction tubes and other vessels. Bovine Serum Albumin can also be used to determine the quantity of other proteins, by comparing an unknown quantity of protein to known amounts of BSA.
Figure S2. Time-course of SDF-1-dependent activation of Erk1/2 and Akt signalling in primary microglial cells. Cultured primary rat microglia were treated for the indicated time with SDF-1 (100 ng/ml) and subsequently analysed for activation (phosphorylation) of (A) Erk1/2 and (B) Akt by Western blotting and the use of phosphospecific antibodies. Protein loading was controlled by additionally staining blots with antibodies recognizing Erk1/2 and Akt independent of their phosphorylation status. Immunoreactive protein bands were quantified by densitometry and corrected for protein loading. Levels of phosphorylated (p)Erk1/2 and pAkt present in untreated controls were set to 1. Bars represent mean ± SD (n = 3). Levels of both pErk1/2 and pAkt were maximal after 3 min of SDF-1 treatment and returned to control levels after 6 min and 10 min respectively. **P , 0.005; *P , 0.02, treatment vs. untreated control. ...
Gang Zhang from the Nilsson group has been investigating the spindle assembly checkpoint in human cells and how it controls the timing of chromosome segregation. In particularly he has focused on understanding how the checkpoint protein Mad1 localized to kinetochores, large protein structures on chromosomes that bind to microtubules. It is known that Mad1 localization is important for checkpoint function but the underlying protein-protein interactions and their regulation have not been uncovered.. In collaboration with the Nielsen lab at CPR the researchers used an in vivo proximity dependent biotinylation approach coupled with mass spectrometry to identify Mad1 interactors. This revealed a so far elusive interaction with the Bub1 checkpoint protein that the researchers further characterized. Interestingly the binding of Mad1 and Bub1 required the phosphorylation of Bub1 by mitotic kinases and a novel generated phospho-specific antibody revealed that Bub1 was specifically phosphorylated on ...
Vasopressin is the primary hormone regulating urine-concentrating ability. Vasopressin phosphorylates the UT-A1 urea transporter in rat inner medullary collecting ducts (IMCDs). To assess the effect of UT-A1 phosphorylation at S486, we developed a phospho-specific antibody to S486-UT-A1 using an 11 amino acid peptide antigen starting from amino acid 482 that bracketed S486 in roughly the center of the sequence. We also developed two stably transfected mIMCD3 cell lines: one expressing wild-type UT-A1 and one expressing a mutated form of UT-A1, S486A/S499A, that is unresponsive to protein kinase A. Forskolin stimulates urea flux in the wild-type UT-A1-mIMCD3 cells but not in the S486A/S499A-UT-A1-mIMCD3 cells. The phospho-S486-UT-A1 antibody identified UT-A1 protein in the wild-type UT-A1-mIMCD3 cells but not in the S486A/S499A-UT-A1-mIMCD3 cells. In rat IMCDs, forskolin increased the abundance of phospho-S486-UT-A1 (measured using the phospho-S486 antibody) and of total UT-A1 phosphorylation ...
The recognition of DNA Double Strand Breaks (DSBs) using a phospho-specific antibody to the histone 2A variant has become the gold standard assay for DNA damage detection. Here we report on the development of the first monoclonal antibody to the phospho-specific form of Drosophila H2AV and characterize the specificity of this antibody to programmed DSBs in oocytes and rereplication sites in endocycling cells by immunofluorescence assays and to DSBs resulting from irradiation in both cell culture and whole tissue by Western blot assays. These studies show that the antibody derived in the study is highly specific for this modification that occurs at DSB sites, and therefore will be a new useful tool within the Drosophila community for the study of DNA damage response, DSB repair, meiotic recombination and chemical agents that cause DNA damage. ...
Human exonuclease 1 (hEXO1) acts directly in diverse DNA processing events, including replication, mismatch repair (MMR), and double strand break repair (DSBR), and it was also recently described to function as damage sensor and apoptosis inducer following DNA damage. In contrast, 14-3-3 proteins are regulatory phosphorserine/threonine binding proteins involved in the control of diverse cellular events, including cell cycle checkpoint and apoptosis signaling. hEXO1 is regulated by post-translation Ser/Thr phosphorylation in a yet not fully clarified manner, but evidently three phosphorylation sites are specifically induced by replication inhibition leading to protein ubiquitination and degradation. We demonstrate direct and robust interaction between hEXO1 and six of the seven 14-3-3 isoforms in vitro, suggestive of a novel protein interaction network between DNA repair and cell cycle control. Binding experiments reveal weak affinity of the more selective isoform 14-3-3σ but both 14-3-3 ...
TY - JOUR. T1 - The properties of the chromosomal architectural HMGB proteins from plants are modulated by differential phosphorylation by protein kinase CK2. AU - Stemmer, C.. AU - Bauw, G.. AU - Fojan, P.. AU - Grasser, K. D.. PY - 2001. Y1 - 2001. M3 - Journal article. VL - 8. SP - 35. JO - Genes, Chromosomes, Genomes. JF - Genes, Chromosomes, Genomes. SN - 0944-6931. ER - ...
RdoA, a serine/threonine kinase, is a member of the Cpx regulon, a stress response pathway, in Salmonella enterica serovar Typhimurium. Phenotypic characterization of rdoA null mutants suggested that RdoA kinase activity affects a wide range of cell functions, which could be the result of both direct and indirect phosphorylation of targets. In a search for RdoAs target(s), the phosphoproteome profile of wild-type and rdoA null S. enterica was examined through phosphoprotein enrichment followed by 2-dimensional gel electrophoresis coupled with phospho-specific fluorescent stains and western blots using phospho-specific antibodies. Three different phosphoprotein enrichment protocols, all based on metal-ion affinity chromatography, were compared for yield and phosphoprotein specificity to determine which would be the most suitable for S. enterica. This study showed that the Phostag Enrich Phosphoprotein kit (PerkinElmer) gave the highest yield, the majority of which were phosphoproteins. These ...
A-Kinase anchoring protein 150 (AKAP150) is required for the phosphorylation of transient receptor potential cation channel subfamily V member 1 (TRPV1) by PKA or PKC in sensory neurons and, hence, affects TRPV1-dependent hyperalgesia under pathological conditions. Recently, we showed that the activation of N-methyl-D-aspartate (NMDA) receptors sensitizes TRPV1 by enhancing serine phosphorylation through PKC in trigeminal nociceptors. In this study, we extended this observation by investigating whether AKAP150 mediates NMDA-induced phosphorylation of TRPV1 via PKC in native sensory neurons in the rat. By adopting a phospho-specific antibody combined with a surface biotinylation assay, we first assessed NMDA-induced changes in the phosphorylation level of serine 800 residues (S800) in TRPV1 delimited to cell surface membrane in cultured trigeminal ganglia (TG). The biotinylation assay yielded that the application of NMDA significantly increased the phosphorylation of S800 (p-S800) of TRPV1 at ...
Regulation of hepatic gluconeogenesis by hormones insulin and glucagon is central to glucose homeostasis. Recent work has proposed that amongst the salt inducible kinase isoforms (SIK1, 2 and 3), members of the AMPK-related kinase family, the SIK2 isoform may play a role as signalling mediator in the control of insulin- and glucagon-regulated hepatic gluconeogenesis. However, the mechanisms of the hormonal-regulation of SIK2 in liver remain controversial, with much of the data based on the studies in non-hepatic tissues/cells. Therefore, the exact molecular regulation of SIK2 by these hormones in the liver required robust and intensive molecular/biochemical research coupled to physiological readout (e.g. gluconeogenesis). My studies with phosphopeptide mapping by mass spectrometry followed by verification with well-characterised phospho-specific antibodies revealed that SIK2 was phosphorylated on Ser343, Ser358, Thr484 and Ser587 in response to glucagon and fasting but not following insulin ...
Mass spectrometry based characterization of differential phosphorylation of signaling proteins by epidermal growth factor and nucleotides. . Download books free in pdf. Online library with books, university works and thousands of documents available to read online and download.
By using signaling pathway- and cell type-specific responses in a combinatorial manner, the immune system signaling network is able to generate a highly diverse set of functional responses despite a limited number of cytokines, cell types, and signaling proteins. In addition to receptors that are coupled to intricate intracellular pathways, specialized cell types express their own set of receptors, and the activation of the same receptor on two different cell types can result in disparate functional responses. This allows sophisticated network behavior despite a limited repertoire of conserved signaling proteins (primarily of the Jak-Stat pathway). Modulating both these levels of the network is the organization of the immune system into distinct compartments, further refining the immune response and ensuring that it is appropriate for the tissue in which the cells resides.. The analysis of immune cell signaling at the network level can be enhanced by approaches that allow cell activities to be ...

antibodies phospho specific Protocols and Video...'antibodies phospho specific' Protocols and Video...

Antibodies, Phospho-Specific: Antibodies directed against immunogen-coupled phosphorylated Peptides corresponding to amino ...
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PLOS ONE: New Phosphospecific Antibody Reveals Isoform-Specific Phosphorylation of CPEB3 ProteinPLOS ONE: New Phosphospecific Antibody Reveals Isoform-Specific Phosphorylation of CPEB3 Protein

Using a new phosphospecific antibody directed to the phosphorylation consensus we found Protein Kinase A and Calcium/Calmodulin ... and describe a robust phosphospecific antibody to study it in future. ... Interestingly, status epilepticus induced by systemic kainate injection in mice led to specific upregulation of the CPEB3 ... and the isoform expression pattern to be cell type-specific. Here, focusing on the alternatively spliced region of CPEB3, we ...
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Phospho-Specific Antibodies | SCBT - Santa Cruz BiotechnologyPhospho-Specific Antibodies | SCBT - Santa Cruz Biotechnology

... serine and threonine in addition to 311 monoclonal antibodies and 1,223 polyclonal antibodies specific for phosphorylated ... Santa Cruz Biotechnology offers antibodies specific for phosphorylated amino acids tyrosine, ... Phospho-Specific Antibodies. Santa Cruz Biotechnology offers antibodies specific for phosphorylated amino acids tyrosine, ... whereas site specific phospho-specific antibodies provide information regarding activation of target proteins at specific ...
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Phospho-Specific Antibodies for Flow Cytometry | Thermo Fisher Scientific - USPhospho-Specific Antibodies for Flow Cytometry | Thermo Fisher Scientific - US

Our phospho-specific antibodies validated for use in flow cytometry are designed for quick and effective assessment of ... Each phospho-specific antibody has been tested in three different flow cytometry buffer systems: IC Fixation and ... Phospho-specific antibody flow cytometric analysis of phosphorylated proteins allows researchers a quick and effective way to ... Testing antibodies in pathway- and cell type-specific experiments helps provide end-users with peace of mind regarding the ...
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Phospho-Specific Antibody | 14 3 3 Antibody | ProSci IncPhospho-Specific Antibody | 14 3 3 Antibody | ProSci Inc

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Phospho-Specific Antibody | 14 3 3 Antibody | ProSci IncPhospho-Specific Antibody | 14 3 3 Antibody | ProSci Inc

ProSci offers primary antibodies for a range of research areas including innate immunity, apoptosis, and more! Order phospho- ... Broad Antibody Catalog , Extensive Antibody Services , In-house USA Labs & Animal Facilities , Established in 1998 ... CUSTOM ANTIBODY SERVICES. * Polyclonal Antibodies * Monoclonal Antibodies * Rabbit Monoclonal Antibody , ProSci Inc ...
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Custom Phospho-Specific Antibody (Poly) Custom Phospho-Specific Antibody (Poly). Custom Phospho-Specific Polyclonal Antibody ... Phospho-peptide antibody purification by 2-steps peptide affinity purification including phospho-peptide column and non-phospho ... Shipment of purified phosphospecific antibody which does not cross-react with. non-phosphospecific peptides (confirmed by ELISA ... Genemed Synthesis offers phosphospecific antibody affinity purification which includes: *Phosphospecific peptide synthesis (up ...
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Phospho-specific antibodies from AMSBIO are affinity-purified rabbit polyclonal or monoclonal antibodies that are monospecific ... AMSBIO has developed a custom phospho-specific antibody production service that is both reliable and produces highly specific ... The purified phospho-specific antibody is fully characterized using both dot-blot and ELISA techniques to ensure product of the ... Phospho-specific antibodies are widely accepted for use in defining regulatory mechanisms that control cell functions such as ...
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Custom Phosphospecific Antibody ProductionCustom Phosphospecific Antibody Production

However, because antibodies specific to the phospho site will represent only a portion of antibodies against the phospho ... Phospho Purification Strategy:. To isolate antibodies specific to the phospho epitope, serum will first be affinity purified ... Purified Antibody Yields:. On average, 2-3 mg of peptide specific antibody will be isolated from affinity purification of 25 ml ... Our Custom Phosphospecific Antibody Production package includes everything needed to obtain affinity-purified antibodies that ...
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Characterization of a Phospho-Specific Antibody for Autophosphorylated GRK1 | IOVS | ARVO JournalsCharacterization of a Phospho-Specific Antibody for Autophosphorylated GRK1 | IOVS | ARVO Journals

Characterization of a Phospho-Specific Antibody for Autophosphorylated GRK1 You will receive an email whenever this article is ... F. S. Chen, C.-K. Chen; Characterization of a Phospho-Specific Antibody for Autophosphorylated GRK1. Invest. Ophthalmol. Vis. ... We sought to develop a phospho-specific antibody for autophosphorylated GRK1 to probe its function under physiological and ... Phospho-specific immunoglobulin (A4102) against autophosphorylated GRK1 was pre-absorbed with nonphosphopeptide and purified ...
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A Mass Spectrometry-based Proteomic Approach for Identification of Serine/Threonine-phosphorylated Proteins by Enrichment with...A Mass Spectrometry-based Proteomic Approach for Identification of Serine/Threonine-phosphorylated Proteins by Enrichment with...

Approach for Identification of Serine/Threonine-phosphorylated Proteins by Enrichment with Phospho-specific Antibodies. ... Approach for Identification of Serine/Threonine-phosphorylated Proteins by Enrichment with Phospho-specific Antibodies ... Anti-FLAG antibody was from Sigma, and anti-filamin-1 antibody was from Santa Cruz Biotechnology (Santa Cruz, CA). ... Approach for Identification of Serine/Threonine-phosphorylated Proteins by Enrichment with Phospho-specific Antibodies ...
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AnaSpec Releases 4 New Phosphospecific Anti-Tau Antibodies - HUM-MOLGENAnaSpec Releases 4 New Phosphospecific Anti-Tau Antibodies - HUM-MOLGEN

Anti-phospho-Tau antibodies are used to identify specific amino acids that are phosphorylated in Tau from normal brains and ... Rabbit anti-Phospho-Tau antibodies were raised against synthetic phosphopeptides. These antibodies were evaluated for ... This represents the industry s largest collection of anti-Tau antibodies - 17 phosphospecific and 9 non-phosphospecific ... Topic: AnaSpec Releases 4 New Phosphospecific Anti-Tau Antibodies anaspec. Member posted 11-21-2008 05:56 PM San Jose, CA ...
more infohttps://hum-molgen.org/bb/Forum1/HTML/000339.html

ECM BIOSCIENCES LLC EB3 (Ser-162), phospho-specific Rabbit Polyclonal Antibody,
 | Fisher ScientificECM BIOSCIENCES LLC EB3 (Ser-162), phospho-specific Rabbit Polyclonal Antibody, | Fisher Scientific

... phospho-specific Rabbit Polyclonal Antibody, 100 μl each, ECM Biosciences EB3 (SER-162) PAB 100UL ... ECM BIOSCIENCES LLC EB3 (Ser-162), phospho-specific Rabbit Polyclonal Antibody, 100 μl each, ECM Biosciences ... EB3 (Ser-162), phospho-specific Rabbit Polyclonal Antibody, 100 μl each, ECM Biosciences ...
more infohttps://www.fishersci.com/shop/products/eb3-ser-162-pab-100ul/501720763

Issues associated with the use of phosphospecific antibodies to localise active and inactive pools of GSK-3 in cells | Biology...Issues associated with the use of phosphospecific antibodies to localise active and inactive pools of GSK-3 in cells | Biology...

... the phosphorylated Tyr279/216 antibodies appear to be specific in western blotting. However, we cannot rule out the ... we examined the specificity of several commercially available anti-phosphorylated GSK-3 antibodies. We show that antibodies ... These antibodies have further been used to determine the subcellular localisation of active and inactive forms of GSK-3, and ... In addition, two antibodies raised to peptides containing the phosphorylated Tyr279/216 epitope recognise an unidentified ...
more infohttps://biologydirect.biomedcentral.com/articles/10.1186/1745-6150-6-4

Development of phospho-specific Rab protein antibodies to monitor in vivo activity of the LRRK2 Parkinsons disease kinase |...Development of phospho-specific Rab protein antibodies to monitor in vivo activity of the LRRK2 Parkinson's disease kinase |...

Polyclonal phospho-specific Rab protein antibodies. We first raised sheep polyclonal antibodies against phospho-peptides ... affinity-purified phospho-specific polyclonal antibodies (all at 1 µg/ml antibody - for only sheep polyclonal antibodies, we ... Sheep polyclonal antibodies. Table 1 summarises the sheep phospho-specific and total Rab polyclonal antibodies generated for ... will complement the selective Rab10 phospho-specific antibodies to assess LRRK2 in vivo activity. Furthermore, the phospho- ...
more infohttp://www.biochemj.org/content/475/1/1

Issues associated with the use of phosphospecific antibodies to localise active and inactive pools of GSK-3 in cells | Biology...Issues associated with the use of phosphospecific antibodies to localise active and inactive pools of GSK-3 in cells | Biology...

... the phosphorylated Tyr279/216 antibodies appear to be specific in western blotting. However, we cannot rule out the ... we examined the specificity of several commercially available anti-phosphorylated GSK-3 antibodies. We show that antibodies ... These antibodies have further been used to determine the subcellular localisation of active and inactive forms of GSK-3, and ... In addition, two antibodies raised to peptides containing the phosphorylated Tyr279/216 epitope recognise an unidentified ...
more infohttps://0-biologydirect-biomedcentral-com.brum.beds.ac.uk/articles/10.1186/1745-6150-6-4

Custom Phospho-Specific antibody Services Affinity antibody - Affinity Biosciences,Cell Signal TransductionCustom Phospho-Specific antibody Services Affinity antibody - Affinity Biosciences,Cell Signal Transduction

Custom Phospho-Specific antibody .Edited in 2014-06-26. Article Category Services Support Orders About Us Contact News antibody ... Affinity-Purified phospho-specific polyclonal antibody production $1199/ project(From Peptide Synthesis to ELISA titer assay). ... antibodies is performed by immuno-affinity purification, using two columns ,which does not cross-react with non-phosphospecific ... Phosphospecific peptide synthesis (up to 15 amino acids)--10 mg used for phosphospecific epitope affinity purification, 10 mg ...
more infohttp://affbiotech.com/article-9-Custom+Phospho-Specific+antibody.html

CRO for rabbit phospho specific PAb:
        Antibody Discussion Forum:
        ABRF Discussion ForumsCRO for rabbit phospho specific PAb: Antibody Discussion Forum: ABRF Discussion Forums

Do any of you have a CRO you trust to make a rabbit PAb specific for a phospho peptide ... purified of course. Frances ... They are very good at the double affinity purification needed to isolate the phospho-specific antibodies. Contact Jordan ... They are very good at the double affinity purification needed to isolate the phospho-specific antibodies. Contact Jordan ... CRO for rabbit phospho specific PAb Hi Frances: We did not make the peptides but the process is that polyclonal antibodies ...
more infohttp://list.abrf.org/groups/antibody/messages/topic/50YaYl8Csgpr5CJJzxEYAU

The autophagy initiator ULK1 sensitizes AMPK to allosteric drugs | Nature CommunicationsThe autophagy initiator ULK1 sensitizes AMPK to allosteric drugs | Nature Communications

To confirm β-isoform specificity of ULK1 we generated a phosphospecific antibody to β2-pSer108 (Supplementary Fig. 4). ... Other β1-AMPK-specific direct activators MT63-7836 and the indole acid derivative PF-0640957737 have shown promise as ... 2b, c). Immunoblot analysis with a phosphospecific antibody confirmed significant increase in pPAK2-Ser141 in S108E-expressing ... Generation of AMPK β2-pSer108 phosphospecific antibody. A phosphorylated synthetic peptide (CSTKIPLIKpSHNDFVAILD, corresponding ...
more infohttps://www.nature.com/articles/s41467-017-00628-y?error=cookies_not_supported&code=070b3ba5-18e6-43d6-8ca3-aa548c15dcdc

Custom Polyclonal Antibody Purification | Thermo Fisher Scientific - USCustom Polyclonal Antibody Purification | Thermo Fisher Scientific - US

Antigen-specific affinity purification Antigen-specific antibody purification is the method of choice for most researchers when ... Phosphospecific Antibody Example. Antiserum is first incubated with the phosphopeptide column and then antibodies are eluted ... This process enables maximum isolation and enrichment of only the most specific antibodies generated to the antigen while ... After a customer confirms the specific reactivity of antibody in corresponding chicken antiserum, eggs collected are pooled and ...
more infohttps://www.thermofisher.com/us/en/home/life-science/antibodies/custom-antibodies/custom-antibody-purification/custom-polyclonal-antibody-purification.html

Casein Kinase 2α Regulates Multidrug Resistance-Associated Protein 1 Function via Phosphorylation of Thr249 | Molecular...Casein Kinase 2α Regulates Multidrug Resistance-Associated Protein 1 Function via Phosphorylation of Thr249 | Molecular...

One rabbit was found to produce MRP1-specific antibodies. Serum containing MRP1-specific antibody was then subjected to ... for phosphospecific antibody) or with 5% nonfat dry milk in Tris-buffered saline/Tween 20, and incubated with primary antibody ... C with primary antibodies, MRP1-Thr249-P (phosphospecific antibody synthesized by Open Biosystems), anti-MRP1 (QCRL-1; Santa ... Next, a phosphospecific antibody to Thr249 (MRP1-Thr249-P) was produced in a rabbit as described under Materials and Methods. ...
more infohttp://molpharm.aspetjournals.org/content/82/3/488.long

ResearchResearch

Generation of phosphospecific antibodies. *Detection of phosphorylation with phosphospecific antibodies (immunoblotting, ... work identified a phosphorylation site in LXRalpha that is important to selectively inhibit the transcription of specific LXR ...
more infohttp://www.ucl.ac.uk/clinical-pharmacology/research

Anti-Cdc25A (phospho S124) antibody (ab182666) | AbcamAnti-Cdc25A (phospho S124) antibody (ab182666) | Abcam

Rabbit polyclonal Cdc25A antibody validated for WB and tested in Human, Mouse and Rat. Immunogen corresponding to synthetic ... ab182666 was purified by affinity-chromatography using epitope-specific phosphopeptide. Non-phospho specific antibodies were ... Primary antibodies. Secondary antibodies. ELISA, Matched Antibody Pairs and Multiplex Immunoassays. Cell and tissue imaging ... All lanes : Anti-Cdc25A (phospho S124) antibody (ab182666) at 1/500 dilution. Lane 1 : UV treated 293 cell extract with ...
more infohttp://www.abcam.com/cdc25a-phospho-s124-antibody-ab182666.html
  • Aalto Bio Reagents its pleased to announce availability of its new mouse monoclonal antibody to Helicobacter pylori flagellin for diagnostic test manufacturers, vaccine developers and researchers globally. (technologynetworks.com)
  • Once antiserum has been selected for purification, it is pooled and the IgG fraction of antibody is isolated by ammonium sulfate precipitation. (thermofisher.com)
  • Antibodies to post-translational modifications such as phosphorylation or acetylation benefit from the use of a multi-column positive and negative purification protocol. (thermofisher.com)
  • After antiserum is incubated in the column, antibodies are eluted using step-wise pH gradient and collected in neutralizing buffer. (thermofisher.com)
  • Antiserum is first incubated with the phosphopeptide column and then antibodies are eluted and subjected to a column containing the non-phosphopeptide. (thermofisher.com)
  • Soluble MHC molecules are commonly used to identify antigen-specific T cells that react to cancers. (biolegend.com)
  • Flex-T™ is BioLegend's technology to study antigen-specific T cells through their TCRs. (biolegend.com)
  • NHEJ is also used to repair DSBs generated during V(D)J recombination when gene regions are rearranged to create the unique antigen binding sites of antibodies and T-cell receptors. (wikipedia.org)
  • Get access to the best antibodies, discovery platforms, and know-how to advance your diagnostic and therapeutic programs. (abcam.com)
  • Custom antibody development and commercial partnerships to advance your diagnostic and therapeutic discovery. (abcam.com)
  • For researchers interested in no-hassle bulk quantities of biofunctional antibodies for mouse injection or human ex vivo studies, GoInVivo™ provides the best solution at exceptional prices. (biolegend.com)
  • Many applications require antibodies to be purified to obtain the best results. (thermofisher.com)
  • Many applications, such as IHC and IF, benefit from this process as antibodies can be used at higher concentrations (lower dilutions) without increasing signal background. (thermofisher.com)
  • Invitrogen eBioscience phospho flow antibodies and related buffer systems are rigorously tested using the methods shown below to help give scientists confidence that they will perform as expected. (thermofisher.com)
  • The recommended buffer system(s) will be noted on the Technical Data Sheet for each specific antibody. (thermofisher.com)
  • Please refer to our table ( Antibody Clone Performance Following Fixation/Permeabilization ) to evaluate the performance of numerous clones in the methanol-based buffer system. (thermofisher.com)
  • A double-stranded gap in DNA will also prevent replication from proceeding, resulting in an incomplete copy of that specific chromosome, targeting the cell for apoptosis. (wikipedia.org)
  • Multiple isoforms of each subunit exist in mammals (α1/2, β1/2, γ1/2/3) and isoform-specific variations in tissue distribution, regulation, and function have been demonstrated. (nature.com)
  • Typically, cells are fixed with formaldehyde to stabilize the cell membrane, and then permeabilized with detergent or alcohol to allow antibodies against intracellular antigens access to stain intracellularly. (thermofisher.com)
  • However, please recognize that surface staining before stimulation can have undesired effects due to activation of the cell caused by antibody binding. (thermofisher.com)