Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
Exclusive legal rights or privileges applied to inventions, plants, etc.
Liquid components of living organisms.
Process of using a rotating machine to generate centrifugal force to separate substances of different densities, remove moisture, or simulate gravitational effects. It employs a large motor-driven apparatus with a long arm, at the end of which human and animal subjects, biological specimens, or equipment can be revolved and rotated at various speeds to study gravitational effects. (From Websters, 10th ed; McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
A novel composition, device, or process, independently conceived de novo or derived from a pre-existing model.
Techniques to partition various components of the cell into SUBCELLULAR FRACTIONS.
Property, such as patents, trademarks, and copyright, that results from creative effort. The Patent and Copyright Clause (Art. 1, Sec. 8, cl. 8) of the United States Constitution provides for promoting the progress of science and useful arts by securing for limited times to authors and inventors, the exclusive right to their respective writings and discoveries. (From Black's Law Dictionary, 5th ed, p1014)
A member of the alkali metals. It has an atomic symbol Cs, atomic number 50, and atomic weight 132.91. Cesium has many industrial applications, including the construction of atomic clocks based on its atomic vibrational frequency.
A plant family of the order Malvales, subclass Dilleniidae, class Magnoliopsida of tropical trees.
A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.
The metabolic process of all living cells (animal and plant) in which oxygen is used to provide a source of energy for the cell.
The third type of glial cell, along with astrocytes and oligodendrocytes (which together form the macroglia). Microglia vary in appearance depending on developmental stage, functional state, and anatomical location; subtype terms include ramified, perivascular, ameboid, resting, and activated. Microglia clearly are capable of phagocytosis and play an important role in a wide spectrum of neuropathologies. They have also been suggested to act in several other roles including in secretion (e.g., of cytokines and neural growth factors), in immunological processing (e.g., antigen presentation), and in central nervous system development and remodeling.
The main structural component of the LIVER. They are specialized EPITHELIAL CELLS that are organized into interconnected plates called lobules.
Messages between computer users via COMPUTER COMMUNICATION NETWORKS. This feature duplicates most of the features of paper mail, such as forwarding, multiple copies, and attachments of images and other file types, but with a speed advantage. The term also refers to an individual message sent in this way.
Transparent, tasteless crystals found in nature as agate, amethyst, chalcedony, cristobalite, flint, sand, QUARTZ, and tridymite. The compound is insoluble in water or acids except hydrofluoric acid.
The study of natural phenomena by observation, measurement, and experimentation.
A technique used to separate particles according to their densities in a continuous density gradient. The sample is usually mixed with a solution of known gradient materials and subjected to centrifugation. Each particle sediments to the position at which the gradient density is equal to its own. The range of the density gradient is usually greater than that of the sample particles. It is used in purifying biological materials such as proteins, nucleic acids, organelles, and cell types.
Relating to the size of solids.
Centrifugation using a rotating chamber of large capacity in which to separate cell organelles by density-gradient centrifugation. (From McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)
The sum of the weight of all the atoms in a molecule.
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.
Lists of words, usually in alphabetical order, giving information about form, pronunciation, etymology, grammar, and meaning.
The amount of mineral per square centimeter of BONE. This is the definition used in clinical practice. Actual bone density would be expressed in grams per milliliter. It is most frequently measured by X-RAY ABSORPTIOMETRY or TOMOGRAPHY, X RAY COMPUTED. Bone density is an important predictor for OSTEOPOROSIS.
Mature LYMPHOCYTES and MONOCYTES transported by the blood to the body's extravascular space. They are morphologically distinguishable from mature granulocytic leukocytes by their large, non-lobed nuclei and lack of coarse, heavily stained cytoplasmic granules.
The soft tissue filling the cavities of bones. Bone marrow exists in two types, yellow and red. Yellow marrow is found in the large cavities of large bones and consists mostly of fat cells and a few primitive blood cells. Red marrow is a hematopoietic tissue and is the site of production of erythrocytes and granular leukocytes. Bone marrow is made up of a framework of connective tissue containing branching fibers with the frame being filled with marrow cells.
White blood cells. These include granular leukocytes (BASOPHILS; EOSINOPHILS; and NEUTROPHILS) as well as non-granular leukocytes (LYMPHOCYTES and MONOCYTES).
Red blood cells. Mature erythrocytes are non-nucleated, biconcave disks containing HEMOGLOBIN whose function is to transport OXYGEN.
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.
Cells contained in the bone marrow including fat cells (see ADIPOCYTES); STROMAL CELLS; MEGAKARYOCYTES; and the immediate precursors of most blood cells.
A species of gram-negative bacteria in the genus ANAPLASMA, family ANAPLASMATACEAE, formerly called Ehrlichia phagocytophila or Ehrlichia equi. This organism is tick-borne (IXODES) and causes disease in horses and sheep. In humans, it causes human granulocytic EHRLICHIOSIS.
A species of gram-negative bacteria and causative agent of severe bovine ANAPLASMOSIS. It is the most pathogenic of the ANAPLASMA species.
A disease of cattle caused by parasitization of the red blood cells by bacteria of the genus ANAPLASMA.
A genus of gram-negative bacteria whose organisms are obligate parasites of vertebrates. Species are transmitted by arthropod vectors with the host range limited to ruminants. Anaplasma marginale is the most pathogenic species and is the causative agent of severe bovine anaplasmosis.
A tick-borne disease characterized by FEVER; HEADACHE; myalgias; ANOREXIA; and occasionally RASH. It is caused by several bacterial species and can produce disease in DOGS; CATTLE; SHEEP; GOATS; HORSES; and humans. The primary species causing human disease are EHRLICHIA CHAFFEENSIS; ANAPLASMA PHAGOCYTOPHILUM; and Ehrlichia ewingii.
Any spaces or cavities within a cell. They may function in digestion, storage, secretion, or excretion.
Blood-sucking acarid parasites of the order Ixodida comprising two families: the softbacked ticks (ARGASIDAE) and hardbacked ticks (IXODIDAE). Ticks are larger than their relatives, the MITES. They penetrate the skin of their host by means of highly specialized, hooked mouth parts and feed on its blood. Ticks attack all groups of terrestrial vertebrates. In humans they are responsible for many TICK-BORNE DISEASES, including the transmission of ROCKY MOUNTAIN SPOTTED FEVER; TULAREMIA; BABESIOSIS; AFRICAN SWINE FEVER; and RELAPSING FEVER. (From Barnes, Invertebrate Zoology, 5th ed, pp543-44)
A 51-amino acid pancreatic hormone that plays a major role in the regulation of glucose metabolism, directly by suppressing endogenous glucose production (GLYCOGENOLYSIS; GLUCONEOGENESIS) and indirectly by suppressing GLUCAGON secretion and LIPOLYSIS. Native insulin is a globular protein comprised of a zinc-coordinated hexamer. Each insulin monomer containing two chains, A (21 residues) and B (30 residues), linked by two disulfide bonds. Insulin is used as a drug to control insulin-dependent diabetes mellitus (DIABETES MELLITUS, TYPE 1).
A primary source of energy for living organisms. It is naturally occurring and is found in fruits and other parts of plants in its free state. It is used therapeutically in fluid and nutrient replacement.
An enzyme that catalyzes the transfer of methyl groups from S-adenosylmethionine to free carboxyl groups of a protein molecule forming methyl esters. EC 2.1.1.-.
A GTP-BINDING PROTEIN involved in regulating a signal transduction pathway that controls assembly of focal adhesions and actin stress fibers. This enzyme was formerly listed as EC 3.6.1.47.
A sub-family of RHO GTP-BINDING PROTEINS that is involved in regulating the organization of cytoskeletal filaments. This enzyme was formerly listed as EC 3.6.1.47.
A large family of MONOMERIC GTP-BINDING PROTEINS that are involved in regulation of actin organization, gene expression and cell cycle progression. This enzyme was formerly listed as EC 3.6.1.47.
Regulatory proteins that act as molecular switches. They control a wide range of biological processes including: receptor signaling, intracellular signal transduction pathways, and protein synthesis. Their activity is regulated by factors that control their ability to bind to and hydrolyze GTP to GDP. EC 3.6.1.-.
Detergent-insoluble CELL MEMBRANE components. They are enriched in SPHINGOLIPIDS and CHOLESTEROL and clustered with glycosyl-phosphatidylinositol (GPI)-anchored proteins.
The principal sterol of all higher animals, distributed in body tissues, especially the brain and spinal cord, and in animal fats and oils.
A tyrosine phosphoprotein that plays an essential role in CAVEOLAE formation. It binds CHOLESTEROL and is involved in LIPIDS transport, membrane traffic, and SIGNAL TRANSDUCTION.
Filamentous proteins that are the main constituent of the thin filaments of muscle fibers. The filaments (known also as filamentous or F-actin) can be dissociated into their globular subunits; each subunit is composed of a single polypeptide 375 amino acids long. This is known as globular or G-actin. In conjunction with MYOSINS, actin is responsible for the contraction and relaxation of muscle.
Endocytic/exocytic CELL MEMBRANE STRUCTURES rich in glycosphingolipids, cholesterol, and lipid-anchored membrane proteins that function in ENDOCYTOSIS (potocytosis), transcytosis, and SIGNAL TRANSDUCTION. Caveolae assume various shapes from open pits to closed vesicles. Caveolar coats are composed of CAVEOLINS.
The main structural proteins of CAVEOLAE. Several distinct genes for caveolins have been identified.
Cyclic GLUCANS consisting of seven (7) glucopyranose units linked by 1,4-glycosidic bonds.

The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis. (1/7983)

Polarized secretion requires proper targeting of secretory vesicles to specific sites on the plasma membrane. Here we report that the exocyst complex plays a key role in vesicle targeting. Sec15p, an exocyst component, can associate with secretory vesicles and interact specifically with the rab GTPase, Sec4p, in its GTP-bound form. A chain of protein-protein interactions leads from Sec4p and Sec15p on the vesicle, through various subunits of the exocyst, to Sec3p, which marks the sites of exocytosis on the plasma membrane. Sec4p may control the assembly of the exocyst. The exocyst may therefore function as a rab effector system for targeted secretion.  (+info)

Studies on a nonpolysomal ribonucleoprotein coding for myosin heavy chains from chick embryonic muscles. (2/7983)

A messenger ribonucleoprotein (mRNP) particle containing the mRNA coding for the myosin heavy chain (MHC mRNA) has been isolated from the postpolysomal fraction of homogenates of 14-day-old chick embryonic muscles. The mRNP sediments in sucrose gradient as 120 S and has a characteristic buoyant density of 1.415 g/cm3, which corresponds to an RNA:protein ratio of 1:3.8. The RNA isolated from the 120 S particle behaved like authentic MHC mRNA purified from chick embryonic muscles with respect to electrophoretic mobility and ability to program the synthesis of myosin heavy chain in a rabbit reticulocyte lysate system as judged by multi-step co-purification of the in vitro products with chick embryonic leg muscle myosin added as carrier. The RNA obtained from the 120 S particle was as effective as purified MHC mRNA in stimulating the synthesis of the complete myosin heavy chains in rabbit reticulocyte lysate under conditions where non-muscle mRNAs had no such effect. Analysis of the protein moieties of the 120 S particle by sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows the presence of seven distinct polypeptides with apparent molecular weights of 44,000, 49,000, 53,000, 81,000, 83,000, and 98,000, whereas typical ribosomal proteins are absent. These results indicate that the 120 S particles are distinct cellular entities unrelated to ribosomes or initiation complexes. The presence of muscle-specific mRNAs as cytoplasmic mRNPs suggests that these particles may be involved in translational control during myogenesis in embryonic muscles.  (+info)

Characterization of nuclear structures containing superhelical DNA. (3/7983)

Structures resembling nuclei but depleted of protein may be released by gently lysing cells in solutions containing non-ionic detergents and high concentrations of salt. These nucleoids sediment in gradients containing intercalating agents in a manner characteristic of DNA that is intact, supercoiled and circular. The concentration of salt present during isolation of human nucleoids affects their protein content. When made in I-95 M NaCl they lack histones and most of the proteins characteristic of chromatin; in 1-0 M NaCl they contain variable amounts of histones. The effects of various treatments on nucleoid integrity were investigated.  (+info)

Isocitrate lyase of Ashbya gossypii--transcriptional regulation and peroxisomal localization. (4/7983)

The isocitrate lyase-encoding gene AgICL1 from the filamentous hemiascomycete Ashbya gossypii was isolated by heterologous complementation of a Saccharomyces cerevisiae icl1d mutant. The open reading frame of 1680 bp encoded a protein of 560 amino acids with a calculated molecular weight of 62584. Disruption of the AgICL1 gene led to complete loss of AgIcl1p activity and inability to grow on oleic acid as sole carbon source. Compartmentation of AgIcl1p in peroxisomes was demonstrated both by Percoll density gradient centrifugation and by immunogold labeling of ultrathin sections using specific antibodies. This fitted with the peroxisomal targeting signal AKL predicted from the C-terminal DNA sequence. Northern blot analysis with mycelium grown on different carbon sources as well as AgICL1 promoter replacement with the constitutive AgTEF promoter revealed a regulation at the transcriptional level. AgICL1 was subject to glucose repression, derepressed by glycerol, partially induced by the C2 compounds ethanol and acetate, and fully induced by soybean oil.  (+info)

Mass spectral study of polymorphism of the apolipoproteins of very low density lipoprotein. (5/7983)

New isoforms of apolipoprotein (apo)C-I and apoC-III have been detected in delipidated fractions from very low density lipoprotein (VLDL) using matrix-assisted laser desorption (MALDI) and electrospray ionization (ESI) mass spectrometry (MS). The cleavage sites of truncated apoC-III isoforms have also been identified. The VLDL fractions were isolated by fixed-angle single-spin ultracentrifugation using a self-generating sucrose density gradient and delipidated using a newly developed C18 solid phase extraction protocol. Fifteen apoC isoforms and apoE were identified in the MALDI spectra and the existence of the more abundant species was verified by ESI-MS. The relative intensities of the apoCs are closely correlated in three normolipidemic subjects. A fourth subject with type V hyperlipidemia exhibited an elevated apoC-III level and a suppressed level of the newly discovered truncated apoC-I isoform. ApoC-II was found to be particularly sensitive to in vitro oxidation. The dynamic range and specificity of the MALDI assay shows that the complete apoC isoform profile and apoE phenotype can be obtained in a single measurement from the delipidated VLDL fraction.  (+info)

Purification and characterization of rat hippocampal CA3-dendritic spines associated with mossy fiber terminals. (6/7983)

We report a revised and improved isolation procedure for CA3-dendritic spines, most of them still in association with mossy fiber terminals resulting in a 7.5-fold enrichment over nuclei and a 29-fold enrichment over myelin. Additionally, red blood cells, medullated fibers, mitochondria and small synaptosomes were significantly depleted. We show by high resolution electron microscopy that this subcellular fraction contains numerous dendritic spines with a rich ultrastructure, e.g. an intact spine apparatus, membranous organelles, free and membrane-bound polyribosomes, endocytic structures and mitochondria. This improved experimental system will allow us to study aspects of post-synaptic functions at the biochemical and molecular level.  (+info)

Two types of HTLV-1 particles are released from MT-2 cells. (7/7983)

The MT-2 cell line transformed by human T-cell leukemia virus type 1 (HTLV-1) contains one complete provirus and seven defective proviruses. Four defective genomes have an identical structure (LTR-MA-deltaCA-pX-LTR) with an open reading frame that spans from MA to pX, giving rise to a 3.4-kb (24S) RNA transcript encoding a chimeric Gag-pX protein, p28. MT-2 cells release two distinct types of virions. The major "classic" type of particle has a buoyant density of 1.155-1.16 g/cm3 and contains the standard HTLV-I structural proteins and reverse transcriptase (RT). In addition, about 5% of particles are "light," approximately 1.12 g/cm3, and contain p28, RT activity, and the 3.4-kb RNA transcript. RT-PCR and in vitro translation indicate that some of the classic HTLV-1 particles package 3.4-kb RNA as well as full-length 8.5-kb RNA. In addition to matrix features, the p28 protein has a motif resembling a zinc finger at the C-terminal, pX0 region, which may play a role in the assembly of the defective light virions.  (+info)

Subcellular localization of oestrogen-induced uterine peroxidase. (8/7983)

The distribution of oestrogen-induced peroxidase in the resuspended 8000g pellet of rat uterine homogenates was examined by centrifugation in a sucrose density gradient. Within 10h of treatment with oestradiol, peroxidase activity was found in a region devoid of catalase or urate oxidase (peroxisomal markers) which did not overlap the fractions containing succinate dehydrogenase (mitochondrial marker) or acid phosphatase (lysosomal marker). The induced uterine enzyme was localized in reticular membrane-bound vesicles with isopycnic density of 1.28g/ml from which it could be released by treatment with detergent.  (+info)

Video articles in JoVE about centrifugation density gradient include Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation, DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation, Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells, A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass Spectrometry, Isolation of Intact Mitochondria from Skeletal Muscle by Differential Centrifugation for High-resolution Respirometry Measurements, Preparation of Liquid-exfoliated Transition Metal Dichalcogenide Nanosheets with Controlled Size and Thickness: A State of the Art Protocol, Isolation of Cellular Lipid Droplets: Two Purification Techniques Starting from Yeast Cells and Human Placentas, Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human
TY - JOUR. T1 - Hydrocarbon molecular markers as indicators of the Late Cenozoic sedimentation on the Amerasian continental margin (Arctic Ocean). AU - Petrova, V. I.. AU - Batova, G. I.. AU - Kursheva, A. V.. AU - Litvinenko, I. V.. AU - Morgunova, I. P.. PY - 2020/4. Y1 - 2020/4. N2 - The main factors controlling the bulk sedimentation in the region of the Mendeleev Rise and the adjacent part of the Arctic Ocean during the late Cenozoic were studied using a complex of geomorphological, lithological, and organic geochemical data. Samples for the study were collected during the cruises of the R/V Akademik Fedorov in 2000, 2005, and 2007 and icebreaker Kapitan Dranitsyn in 2012. Analysis of the group and molecular compositions of the dispersed organic matter (DOM) in bottom sediments has shown that the input of terrigenous sediments enriched with abrasion products of lithified rocks from the eastern source province determines the Holocene-Pleistocene sedimentation on the continental slope of the ...
An investigation of the distribution of immunoglobulins in the external secretions of the hamster revealed the predominance of γA. Only γA was found in saliva, whereas colostrum and urine both contained γ2 in addition to γA. Hamster γA was present in the serum at concentrations that were insufficient to allow its visualization by routine immunoelectrophoresis. No differences in electrophoretic mobility were found between serum and secretory γA regardless of derivation. The secretory γA from saliva, colostrum and urine had sedimentation characteristics intermediate between serum γA and 19S markers in sucrose density gradient ultracentrifugation studies. Similar results were obtained on Sephadex G-200 gel filtration. Isolated secretory γA had an s020,w of 15.2S. Mild reductive procedures did not appear to alter the sedimentation characteristics of this molecule. No antigenic differences were found between serum, salivary, colostral and urinary γA.. ...
Sigma-Aldrich offers Sigma-D1556, OptiPrep™ Density Gradient Medium for your research needs. Find product specific information including CAS, MSDS, protocols and references.
AXIS-SHIELD Density Gradient Media - Macromolecule indexes - Isolation of cells, cell organelles, subcellular membranes, viruses and macromolecules using centrifugation techniques
mRNAs surrounded by polysomes are ready for translation into proteins (Warner et al., 1963); these mRNAs are defined as polysomal-mRNAs (Mustroph et al., 2009). The process is affected by various growth conditions or surrounding situations. Microarray analysis is a powerful tool for detecting genome-wide gene expression. Therefore, using polysomal-mRNAs for microarray analysis can reflect the gene translation information (the translatome) under different developmental stages or environmental conditions from eukaryotes. Polysomal-mRNAs can be collected from the polysomal fraction by sucrose-gradient separation for further quantitative PCR or microarray assay. We modified a protocol (Mustroph et al., 2009) for collecting polysomal-mRNAs via sucrose-gradient separation to eliminate monosomal-mRNA contamination from pLAT52:HF:RPL18 Arabidopsis. This transgenic Arabidopsis uses a pollen-specific promoter (ProLAT52) to generate epitope-tagged polysomal-RNA complexes that could be purified with a specific
Transcriptional regulation has been the main focus for gene regulation in the past. However, a tremendous amount of evidence from recent studies also indicates that translational regulation plays a key role during development, cell cycle control, and mechanisms related to acute drug resistance. Gene expression analysis on actively translated mRNA transcripts provides a unique approach to study post transcriptional regulation. Previous studies have relied on a traditional sucrose gradient ultracentrifugation procedure to isolate polysome complexes and requires a large amount of cells (up to 500 million cells). As a result, this still remains a major bottleneck for the investigation of post transcriptional regulation with limited quantities of clinical samples. Therefore, there is an urgent need to develop a novel approach to isolate actively translated polysomes from a small number of cells (10 to 500 cells). The new approach will allow us to systematically study potential translational ...
Blinded analyses of the concentrations of binding proteins for retinol and retinoic acid (CRABP) in homogenates of cancer and normal tissue aliquots obtained from human cervix, endometrium, ovary, breast, and lung were carried out by the sucrose gradient ultracentrifugation technique. In carcinomas of the cervix and endometrium, CRABP mean values of 50.4 and 123.2 pmol/g tissue, respectively, were detected. Such concentrations represent a 3- and 4-fold increase over the mean values of CRABP in the normal cervix (16.9 pmol/g) and normal endometrium (30.8 pmol/g), respectively. In carcinomas of the ovary, the mean CRABP level was 128.6 pmol/g compared to a maximal mean value of ≦0.46 pmol/g in the normal ovary. Elevated levels of CRABP were also found in breast and lung carcinomas compared to the amounts detected in the same patient in normal tissue aliquots of the same organ.. The differences between CRABP concentrations in cervical, endometrial, ovarian, and breast carcinomas and those in ...
Specialized tube for rapid PBMC isolation by density gradient centrifugation. Manufactured under cGMP and can be used for IVD applications.
In order to study the release of ATP from a neuronal preparation thought to be purinergic, isolated varicosities were prepared from myenteric plexus by mincing and homogenizing the longitudinal muscle of guinea pig ileum in 0.32 M sucrose. The presence of varicosities within the crude preparation (P2), isolated by differential centrifugation, was confirmed by electron microscopy and by the presence of occluded lactate dehydrogenase as a cytoplasmic marker. Varicosities were purified further from the P2 fraction on a discontinuous sucrose density gradient and characterized morphologically. Varicosities resembling cholinergic, purinergic, and adrenergic axonal terminals were identified. The release of ATP from both crude and purified varicosities was detected by monitoring the light produced when the released ATP reacted with firefly luciferin-luciferase which was present in the incubation medium. Elevated extracellular K+ or Rb+ caused the release of ATP, whereas elevated Na+ and Li+ did not. ...
Assays detecting antigen (Ag)-specific T-cell responses in immune-mediated processes are increasingly employed to understand disease pathogenesis and immune staging. The quantity and quality of starting peripheral blood mononuclear cell (PBMC) preparations are important factors in the performance of such assays. We therefore compared final PBMC yield and function by modifying parameters at the blood drawing, storage and processing steps. While drawing blood in vacuum-driven tubes or syringes and separating PBMCs on density gradients using standard or membrane (Leucosep (R)) tubes made no difference, storing tubes for 18 h without any agitation led to PBMC preparations contaminated with granulocytes and decreased interferon (IFN)-gamma enzyme-linked immunospot (ELISpot) responses. Even agitated blood showed a trend towards reduced ELISpot responses and increased human leukocyte Ag (HLA) multimer readouts when stored for 18 h compared to 3 h. These changes were reduced by diluting blood prior to ...
Constitutive secretory vesicles carrying heparan sulfate proteoglycan (HSPG) were identified in isolated rat hepatocytes by pulse-chase experiments with [35S]sulfate and purified by velocity-controlled sucrose gradient centrifugation followed by equilibrium density centrifugation in Nycodenz. Using this procedure, the vesicles were separated from plasma membranes, Golgi, trans-Golgi network (TGN), ER, endosomes, lysosomes, transcytotic vesicles, and mitochondria. The diameter of these vesicles was approximately 100-200 nm as determined by electron microscopy. A typical coat structure as described for intra-Golgi transport vesicles or clathrin-coated vesicles could not be seen, and the vesicles were not associated with the coat protein beta-COP. Furthermore, the vesicles appear to represent a low density compartment (1.05-1.06 g/ml). Other constitutively secreted proteins (rat serum albumin, apolipoprotein E, and fibrinogen) could not be detected in purified HSPG-carrying vesicles, but banded in ...
The dynamics of the flow inside an evaporating sessile droplet of water with polystyrene micro-spheres of 1.0 μm in diameter in suspension is described. The initial volume of the droplets is in the ra
Separation of particles according to density by employing a gradient of varying densities. At equilibrium each particle settles in the gradient at a point equal to its density. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed ...
Cytochrome P-450 in rat liver nuclear membranes of untreated, PB-pretreated, or 3-MC-pretreated animals was spectrally similar to corresponding liver microsomes. Epoxide hydrase activity of liver nuclear membrane and microsomes was induced by PB administration; 3-MC did not cause any induction. The same results were obtained when either the DNase digestion method or sucrose density centrifugation technique for preparation of nuclear membranes was used.. ...
Total CD4+ Th cells from IRF4+/+ or −/− mice were purified using the first steps of the multisort kit (Miltenyi Biotec) as described previously (5). 106/ml of purified Th cells were stimulated in vitro for 72 h with 5 μg/ml of immobilized anti-CD3 mAb, 2.5 μg/ml of soluble anti-CD28 mAb (BD Biosciences), or 100 U/ml of recombinant human IL-2 as described previously (5). After a resting period of 48-72 h in the absence of anti-CD3 mAb, but in the presence of IL-2, viable cells were purified on a Ficoll density gradient and restimulated in the presence of IL-2 either with anti-CD3 (immobilized at 5 μg/ml) or with 1 μg/ml of soluble anti-Fas mAb Jo2 (Becton Dickinson) together with 2 μg/ml of protein G (Sigma-Aldrich). Apoptosis was analyzed by annexin V and propidium iodide (PI) staining as described previously (12) at the indicated times after secondary stimulation. Viable as well as dead cells were included in this analysis according to forward and side scatter characteristics. The role ...
Equilibrium sedimentation uses a gradient of a solution to separate particles based on their individual densities (mass/volume). A pivotal aspect about this type of sedimentation is that it is completely independent of the shape of the molecule. It is used to purify the differential centrifugation. A solution is prepared with the densest portion of the gradient at the bottom. Particles to be separated are then added to the gradient and centrifuged. Each particle proceeds until it reaches an environment of comparable density. Such a density gradient may be continuous or prepared in an incremental fashion. For instance, when using sucrose to prepare density gradients, one can carefully float a solution of 40% sucrose onto a layer of 45% sucrose and add further less dense layers above. The homogenate, prepared in a dilute buffer and centrifuged briefly to remove tissue and unbroken cells, is then layered on top. After centrifugation typically for an hour at about 100,000 x g, disks of cellular ...
We investigated the function of Cav1. 4 MgCl2. The intracellular alternative included (in millimeter focus) 130 beliefs < .05 were considered significant. Outcomes Sucrose-density lean fractionation of protein involved in depolarization-induced insulin release in Inches-1 Cav1 and cells.2/II-III cells The KATP funnel, made up of Kir6.2 and SUR1 subunits, has a central function in the insulin release stimulated by blood sugar and sulfonylureas. The localization was examined by us of Kir6.2, SUR1, and EPAC2 in lipid rafts by fractionating the Triton A100-insoluble part of Cav1 and Inches-1.2/II-III cell lysates in discontinuous sucrose gradients. EPAC2 is normally reported to interact straight with both Piccolo (21) and SUR1 (19), and we discovered that both EPAC2 and SUR1 are extremely focused in lipid number fractions of sucrose gradients (the user interface of 5% and 30% sucrose) in both Inches-1 cells and Cav1.2/II-III cells (Figure 1). We assessed the localization of the KATP funnel subunit ...
We investigated the function of Cav1. 4 MgCl2. The intracellular alternative included (in millimeter focus) 130 beliefs < .05 were considered significant. Outcomes Sucrose-density lean fractionation of protein involved in depolarization-induced insulin release in Inches-1 Cav1 and cells.2/II-III cells The KATP funnel, made up of Kir6.2 and SUR1 subunits, has a central function in the insulin release stimulated by blood sugar and sulfonylureas. The localization was examined by us of Kir6.2, SUR1, and EPAC2 in lipid rafts by fractionating the Triton A100-insoluble part of Cav1 and Inches-1.2/II-III cell lysates in discontinuous sucrose gradients. EPAC2 is normally reported to interact straight with both Piccolo (21) and SUR1 (19), and we discovered that both EPAC2 and SUR1 are extremely focused in lipid number fractions of sucrose gradients (the user interface of 5% and 30% sucrose) in both Inches-1 cells and Cav1.2/II-III cells (Figure 1). We assessed the localization of the KATP funnel subunit ...
Results. Western blot analysis of PKCα and PKCγ translocation. N/N 1003A cells were treated with 200 nM TPA (a conventional PKC activator) for 60 min, 10 ng/ml EGF, or 15 ng/ml bFGF. Confluent cells were taken up in ice cold 50 mM Tris, 20 mM MgCl2, 25 μg/ml aprotinin, and 25 μg/ml leupeptin, pH 7.5 and sonicated. The lysate was spun down for 60 min at 35,000 rpm (100,000x g). Western blot analysis was performed on the pellet that contained plasma membrane and the supernatant that contained cytosolic components.. Upon activation by TPA, PKCα and PKCγ should translocate from the cytosol to the plasma membrane [25]. In order to determine if PKCα and PKCγ translocate to the plasma membrane in N/N 1003A cells upon activation, TPA was added to the cell culture media for 60 min. PKCα did translocate to the membrane (pellet fraction) upon activation of the enzyme by TPA (Figure 1A, lane 2 versus lane 4). PKCγ also translocated to the membrane (pellet fraction, Figure 1B, lane 2 versus lane ...
Powered by Pure, Scopus & Elsevier Fingerprint Engine™ © 2021 Elsevier B.V We use cookies to help provide and enhance our service and tailor content. By continuing you agree to the use of cookies. ...
The present invention relates to methods of enriching for desired cell population from cell sources, such as body fluids, dispersed tissue specimens and cultured cells. In particular, the present inve
As of today, there are 39 hydrogen fueling stations in the U.S., all but four of them in the state of California. They serve roughly 39 million Californians, whereas 284 million Americans outside the state have no access to hydrogen fuel-cell cars nor stations at which to refill them. Germany, on the other hand, has 83 million residents-and 45...
Sucrose Density Gradient Centrifugation Separation of Gold - Sensor basics: Types, functions and applications Martin Rowe while the application of sensors and their associated algorithms has multiplied and changed the
Gradient Makers, C.B.S. Scientific - Each (10 ML) : Designed for use in sucrose density gradient centrifugation, gradient gel electrophoresis, and li
Sucrose density gradient centrifugation& recover fractions-,SDS-PAGE(10% gel) (GM1 would migrate with the dye front)-,electro-transfer to nitrocellulose (or PVDF) membrane -,blocking with 4% skimmilk for 30min-, incubate with Ctx-HRP in 4% skimmilk for 1h-,wash PBS-Tween x 3times-,Detection with ECL. ...
Gravity currents impact significantly the environment and humans life. Understanding their internal structure and their dynamics is fundamental for modelling purposes. In this study we observe experimentally how bed porosity affects the dynamics of density currents. We will focus on density and velocity profiles of currents traveling over porous bed, sinking and entraining water from its cavities. This abstract presents the preliminary results of this study which are relative to density currents travelling over the reference substratum, a smooth impermeable bed. The tests presented will be used as a reference to observe how the water entrained from the bottom cavities can change the structure of the current and are herein used to validate the experimental techniques. Brine water is injected at constant discharge in a 7.5 m long, 0.3 m wide and 0.3 m deep flume while instantaneous velocity and density profiles are simultaneously acquired 3 m downstream of the inlet. Initial excess densities and inlet
pluriSelect separation tubes have been developed for optimal separation of leukocytes and peripheral blood mononuclear cells (PBMC) from whole blood and bone marrow. pluriMate and TwinSpin prevents you from time-consuming and laborious overlaying of the sample material. During centrifugation, Leukocytes, lymphocytes and PBMCs are separated from unwanted erythrocytes and granulocytes, depending on the density gradient media ...
Find out all of the information about the Deltalab product: sedimentation analysis microplate 141746. Contact a supplier or the parent company directly to get a quote or to find out a price or your closest point of sale.
try using percol gradient centrifugation. ive forgotten the manufacturer. they do have protocols for separating live/dead cells. you will have to optimize the percol density(s) for your type of cells. you may also have to use more than one density gradient of percol to separate the layers of dead and live cells.good luck. john. ...
Adenovirus are efficient gene delivery systems. The standard method for purification of adenoviral vectors is based on using a cesium chloride (CsCl) density gradient combined with ultracentrifugation. This method is suitable for small-scale purification and is less expensive than column chromatography or commercial purification kits.
Studies on the composition and structure of plant viruses can only be attempted after their purification from infected tissue homogenates. Owing to their intrinsic differences, however, the ease with which different viruses can be purified varies considerably. A few relatively stable viruses, e.g. tobacco mosaic virus or turnip yellow mosaic virus, can be treated with high concentrations of salt and precipitated by acid or alcohol without inactivation. With less stable viruses such methods are usually unsuccessful. The particles of such viruses, however, can be sedimented by ultracentrifugation and procedures developed frequently involve two treatments: (a) differential centrifugation and concentration after initial clarification of buffered homogenates such as with organic solvents (Steere, 1956; Tomlinson, Shepherd & Walker, 1959; Wetter, 1960), (b) further fractionation by rate or equilibrium density gradient centrifugation as developed by Brakke (1960). Because of the small capacity of the
Technetium-99m-Exametazime (HMPAO) is widely used for radiolabelling leucocytes for localization of infection. The subcellular distribution of radionuclide in the labelled cells and the distribution of radioactivity among the leucocyte population are incompletely understood. Frozen section autoradiography was used to determine quantitatively the distribution of Tc-99m in leucocytes labelled with Tc-99m-Exametazime. Sections of rapidly frozen suspensions of labelled leucocytes in plasma were autoradiographed on Ilford K2 emulsion and stained with haematoxylin and eosin. Neutrophils, eosinophils and mononuclear cells were separated by Percoll density gradient centrifugation. Cell nuclei were isolated by a rapid cell-breakage and fractionation method. In a typical experiment mean grain densities [grains/100 mu m(2) (ESD)] over cells were: eosinophils 31.2 (18.4), neutrophils 3.5 (3.5), mononuclear cells 4.2 (5.1). Mean grain numbers per cell (ESD) were: eosinophils 13 (6.8), neutrophils 1.3 (1.3), ...
The molecular nature and replicative behavior of R factor 222 was examined in Proteus mirabilis . In deoxyribonucleic acid (DNA} from R+ P. mirabilis , R factor 222 was identified by CsCl density gradient centrifugation as 2 satellite DNA bands at densities corresponding to 50 and 58 moles percent guanine plus cytosine (% GC) . Replication of the 50 and 58% GC components of R factor 222 in P. mirabilis was analyzed during growth in the presence and absence of chloramphenicol (CAM} and after shifting exponentialand stationary- phase cells to conditions which inhibit host protein or DNA synthesis . CAM reduced the cellular growth rate but increased the amount of both R factor components relative to host chromosomal DNA . However, the 58% GC component showed a larger proportionate increase. This was inferred to indicate reduced synthesis of an inhibitor that acts on both R factor components and an initiator required for replication of the 50% GC component . Replicative patterns observed after shifting
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
On Wed, 30 Sep 1998, Patrick wrote: , Hi all. , , A request from first-time user of CsCl/EtBr for plant DNA purification: , Upon addition of ethidium bromide to my phenol/chloroform purified , Plant-DNA solution in CsCl, I observe a change of color from normal , orange-red to light cherry red. If I remember correctly this was also the case, when I have purified DNA with CsCl density gradient centrifugation. , Check http://www.biologie.uni-ulm.de/bio2/knoop/images/ethidium.jpg , for visual impression... Unfortunatly my display doesnt show the correct colours. , After centrifugation color accumulated COMPLETELY with a sluggish , material on top of the gradient. Considering the possibility that some , remaining protein/polysaccharides may be responsible for this, I have , taken the remainder of clear solution and repeated the EtBr addition. No , improvement - same color change (even after repeating this for the 4th , time!) , , Anybody ideas what is going on? , I dont know, what is going on, but ...
Abstract: t had been reported that the GCHV(Gtass Carp Hemorrhage Virus)is a new member.in Reoviri-dae and has 11 polypeptides by our laboratory,We did further experiments on the viral polypeptides.The virions(GCHV-875)were purified from the medium of infection culture cells by CsCl density gradient centrifugation,dissolved in the buffer containing Tris-HCl 250mmol/L,glycerin 25%,SDS86mmol/L,2-ME 320mmol/L(pH6.9)and then heated at 100℃for 3 min.After that,by addingan equal volume of FITC solution(2mg/M)in 1 mol/L NaHCO3,the viral polypeptides were labelledwith FITC,and analysed by SDS-PAGE. All the viral structural polypeptides(11 bands),namelyVP130,VP120,VP113,VP107,VP75,VP65,VP60,VP52,VP42,VP39 and VP31,wete visible under. UV light.These polypeptides were retrieved by cutting down each band from the gel and demonstrated kept immunological property.The method can determine 500ng protein band in gel under UVlight.. ...
A simple technique is described for the isolation of Toxoplasma gondii tissue cysts from fetal ovine brain by centrifugation on a discontinuous density gradient of 30 per cent and 90 per cent Percoll (colloidal silica solution). Brain samples from 51 aborted ovine fetuses were examined by both the Percoll and mouse inoculation techniques; eight infections were detected by the Percoll technique compared to 12 by mouse inoculation. Possible reasons for this discrepancy and the scope for improving the Percoll technique are discussed.. ...
An analytical ultracentrifluge was used to characterize the behaviors of ultracentrifugal sedimentation of bovine serum albumin solutions. We obtained data both for the sedimentation velocity and for the growth rate of the sediment that accumulates during ultracentrifugation. Effects of the rotor speed, the solution concentration, and the volume of the solution contained in the cell on the sedimentation behaviors were investigated experimentally. The sedimentation behaviors and the properties of the compressible sediment were explained well in terms of the theory which has been applied with respect to particulate suspension. Finally, it was shown that such factors of the solution environment as the solution pH and the salt concentration have a large effect on the sedimentation behaviors.. ...
Mitochondria are essential for the manufacturing of major and secondary metabolites, which largely decide the standard of fruit. Nevertheless, a technique for isolating high-quality mitochondria is at present not out there in citrus fruit, stopping high-throughput characterization of mitochondrial capabilities. Right here, primarily based on differential and discontinuous Percoll density gradient centrifugation, we devised.... ...
Mitochondria are essential for the manufacturing of major and secondary metabolites, which largely decide the standard of fruit. Nevertheless, a technique for isolating high-quality mitochondria is at present not out there in citrus fruit, stopping high-throughput characterization of mitochondrial capabilities. Right here, primarily based on differential and discontinuous Percoll density gradient centrifugation, we devised.... ...
Outer-arm dynein purified from trout spermatozoa was disrupted by low-ionic-strength dialysis, and the resulting subunits were separated by sucrose density-gradient centrifugation. The intact 19 S dynein, containing the alpha- an beta-heavy chains, intermediate chains (ICs) 1-5 and light chains (LCs) 1-6, yielded several discrete particles: a 17.5 S adenosine triphosphatase (ATPase) composed of the alpha- and beta-chains ICs 3-5 and LC 1; a 9.5 S complex containing ICs 1 and 2 together with LCs 2, 3, 4, and 6; and a single light chain (LC 5), which sedimented at approximately 4 S. In some experiments, ICs 3-5 also separated from the heavy chain complex and were obtained as a distinct subunit. Further dissociation of the 17.5 S particle yielded a 13.1 S ATPase that contained the beta-heavy chain and ICs 3-5. The polypeptide compositions of the complexes provide new information on the intermolecular associations that occur within dynein. Substructural features of the trout dynein polypeptides also were
Harvest cowpea plants about 2 weeks after inoculation, then homogenize at 4°C in two volumes of 0.5 M citrate buffer (pH 7.0) containing 0.1% thioglycollic acid. Express juice through cheesecloth, and add 20 ml carbon tetrachloride to every 100 ml extract. Shake the extract for 15 min, and clarify by low-speed centrifugation. Concentrate the virus by three cycles of differential centrifugation. Resuspend the pellets from high speed centrifugation in 0.01 M citrate buffer. Purify further by sucrose density-gradient centrifugation (Tsuchizaki et al., 1971).. ...
Exosomes purification and analyses comprise a fast evolving research area; more than 70% of published research on exosomes has been done within the last six years. Challenges to researchers working with exosomes include setting up density gradients by hand, because it is tedious, time-consuming and subject to user, lab, and method variability.
Previous studies in this laboratory have allowed the formulation of a model for the molecular arrangement of C5, C6, C7, C8, and C9 on the surface of cells undergoing immune cytolysis with an assigned cumulative m.w. of 995,000. To verify directly the existence of a C5-C9 complex, serum samples containing radiolabeled terminal components were activated at 37°C with EA, antigen-antibody complexes, CVF, inulin or zymosan. Subsequent sucrose density gradient ultracentrifugation showed that all treatments cited led to the formation, in varying degrees, of rapidly sedimenting material which incorporated C5, C6, C7, C8, and C9, but not C3. The reaction was inhibited by 0.01 M EDTA and 0°C. The complex had a sedimentation coefficient of 22.4S, a diffusion coefficient of 1.98 × 10-7 cm2 sec-1 and thus a calculated m.w. of 1.04 × 106.. ...
Purification of mitochondria and mitochondrial subfractionation. Mitochondria were purified from brain tissue using the discontinuous sucrose gradient method. Briefly, brain homogenate was made in ice-cold homo-buffer (0.32 M sucrose, 20 mM Tris-HCl, pH 7.4) and spun at 900 g, 4°C, for 10 minutes. The supernatant was transferred to another clean tube and spun at 10,000 g, 4°C, for 10 minutes. The resultant pellet, enriched for mitochondria, was resuspended in 2 ml homo-buffer, loaded on top of a sucrose gradient (1.2 M, 0.8 M, and 0.32 M sucrose; 20 mM Tris-HCl, pH 7.4) and spun at 53,000 g, 4°C, for 2 hours. The white band at the interface between medium (0.8 M) and heavy (1.2 M) solutions was collected as highly purified mitochondria. Mitochondria from cultured cells were isolated using a kit (catalog no. 89874) from Pierce. Mitochondrial subfractionation was carried out as described by Hovius et al. (40). Briefly, purified mitochondria (1 mg) were resuspended in 500 μl ice-cold buffer (10 ...
Cells. The Raji control cell line and the cell lines expressing either DC-SIGN (Raji-DC-SIGN) or L-SIGN (Raji-L-SIGN) were cultured as previously described (4). PBMCs were isolated from buffy coats by standard Ficoll-Hypaque density centrifugation, activated with phytohemagglutinin (3 μg/ml), and cultured in RPMI medium containing 10% FCS, penicillin (100 units/ml), and streptomycin (100 units/ml). On day 3 the cells underwent CD4+ enrichment by incubation with CD8 immunomagnetic beads (Dynal Biotech) and were negatively selected according to the manufacturers instructions and cultured with IL-2 (100 U/ml). DCs for the single-cycle transmission assay were generated from fresh PBMCs with cells layered on a standard Percoll gradient (Amersham Pharmacia). The light fraction with predominantly monocytes was collected, washed, and seeded in 24-well or 6-well culture plates at a density of 5 × 105 cells or 2.5 × 106 per well, respectively. After 60 minutes at 37°C, the adherent cells were ...
Cells. The Raji control cell line and the cell lines expressing either DC-SIGN (Raji-DC-SIGN) or L-SIGN (Raji-L-SIGN) were cultured as previously described (4). PBMCs were isolated from buffy coats by standard Ficoll-Hypaque density centrifugation, activated with phytohemagglutinin (3 μg/ml), and cultured in RPMI medium containing 10% FCS, penicillin (100 units/ml), and streptomycin (100 units/ml). On day 3 the cells underwent CD4+ enrichment by incubation with CD8 immunomagnetic beads (Dynal Biotech) and were negatively selected according to the manufacturers instructions and cultured with IL-2 (100 U/ml). DCs for the single-cycle transmission assay were generated from fresh PBMCs with cells layered on a standard Percoll gradient (Amersham Pharmacia). The light fraction with predominantly monocytes was collected, washed, and seeded in 24-well or 6-well culture plates at a density of 5 × 105 cells or 2.5 × 106 per well, respectively. After 60 minutes at 37°C, the adherent cells were ...
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
Centrifugation is the use of the centrifugal forces generated in a spinning rotor to separate biological particles, such as cells, viruses, sub‐cellular organelles, macromolecules (principally proteins and nucleic acids) and macromolecular complexes (such as ribonucleoproteins and lipoproteins)
The growth of a mouse leukemia virus in an established mouse cell line was examined after the line became contaminated with an unidentified Mycoplasma species. The contaminated cultures grew well in small plastic cultures dishes, but they could not be propagated in larger roller bottles unless the growth medium was changed frequently. Cells from Mycoplasma-contaminated and Mycoplasma-free cultures were exposed to 3H-labeled uridine for 24 hr. Culture fluids were harvested 2 or 24 hr after labeling and purified by centrifugation through discontinuous sucrose gradients. Considerably less uridine-3H-labeled virus was recovered from supernatant fluids of Mycoplasma-contaminated cultures than from Mycoplasma-free cultures. Equilibrium sedimentation in sucrose gradients of uridine-3H-labeled material from culture supernatants of contaminated cultures produced 3H peaks at buoyant densities of 1.20 to 1.24 and 1.16 to 1.18 g/ml. Virus titers in culture fluids from Mycoplasma-contaminated cultures were ...
The present invention generally encompasses the control of the release rate of agents from a polymeric matrix. This control over the release rate of agents provides for control over, inter alia, the therapeutic, prophylactic, diagnostic, and ameliorative effects that are realized by a patient in need of such treatment. In addition, the control of the release rate of agents also has an effect upon the mechanical integrity of the polymeric matrix, as well as a relationship to a subjects absorption rate of the absorbable polymers.
Ns by differential centrifugation. B and C. Immunoblot analysis of soluble/ insoluble fractions separated by differential centrifugation. FKIPS DCARD stable
Plasma membrane(PM) protein accounts for a small fraction of total cellular protein in plants but performs a very critical role in plant physiology. Isolation and purification of PM protein from plant tissues have been traditionally done by sucrose density ultracentrifugation and aqueous two-phase partitioning. These methods, while relatively effective, require ultracentrifugation and large amount of starting material. The procedures are usually tedious and time consuming.To overcome the shortcomings, we have developed this PM isolation kit. Plant tissues are first sensitized by buffer A, homogenized, and pass through a specialized filter cartridge that allows homogenates to pass through with a zigzag path. The cell membranes are ruptured into a range of predefined size during the process. Native plasma membranes are separated from a mixture of un-ruptured cells, nuclei, cytosol and organelles by subsequent differential centrifugation and density centrifugation without using ...
I am having trouble isolating mtDNA using CsCl gradients. Is there an easier way to isolate mtDNA? I was wondering if anyone could give me some help! This is becoming a really big problem!!!!! Thank oyu very much, Rodney Earl Pettway Rodney Earl Pettway Department of Plant Pathology and Microbiology Texas A&M University College Station, TX 77843 Pettway at ppserver.tamu.edu ...
Density marker beads are small colored microspheres of known density that are used for calibrating density gradients and determining density in gradient columns
StraightFrom® Whole Blood MicroBeads for cell isolation directly from whole blood without the need for density gradient centrifugation. - 日本
Find a potocol and data for the isolation of exosomes without density gradient centrifugation and fast screening of isolated exosomes by flow cytometry. | Ireland
JML-135-2007-105-114. This paper contains the results of a new experimental study of the effect of temperature on density, refractive index on mixing and ultrasonic velocity for a number of linear n-alkanes conbined with ethanol. A perusal of deviations between the experimental and calculated derived magnitudes shows that the predictive procedures give a qualitative estimation for the studied mixtures due to their high nonideality.. ...
2014.1-2015.12 Academy of Mathematics & Systems Science Postdoc within the research center NCMIS, working with Prof. Ping Zhang on density patch problem ...
View Notes - ps_2 from CHEMICAL E 20.410j at MIT. DOWNSTREAM PROCESSING Problem Set #2 Problem 1 A key to understanding centrifugation is understanding the equations that describe it. One can
The classical procedure for estimating the approximate size of a protein is by its sedimentation coefficient, determined either using an analytical ultr...
ca ---------- -cuc a u gu a cu 5 gugcu ggggu aggag cag gcu cggg cgu c ,,,,, ,,,,, ,,,,, ,,, ,,, ,,,, ,,, c 3 cacgg uccca uccuc guc cgg gucc gua a gga aacucacucu cucc - u -- g cc ...
Carr, Dr Steven M. "CsCl density-gradient centrifugation". www.mun.ca. Retrieved 2017-04-03. Martinez, Lluis. "Magnetic DNA ... With high speed buoyant density ultracentrifugation, a density gradient is created with caesium chloride in water. DNA will go ... to the density that reflects its own, and ethidium bromide is then added to enhance the visuals the nucleic acid band provides ...
Timonen T, Saksela E (1980). "Isolation of human NK cells by density gradient centrifugation". Journal of Immunological Methods ... The demonstration that density gradient-isolated large granular lymphocytes were responsible for human NK activity, made by ... Using discontinuous density centrifugation, and later monoclonal antibodies, natural killing ability was mapped to the subset ...
Forster, Mark; Flenley, John R. (1993). "Pollen purification and fractionation by equilibrium density gradient centrifugation ...
OptiPrep™: The ideal density gradient medium, Axis-Shield Density Gradient Media. Accessed on line Nov. 19, 2015. Waite J.A, ... Post centrifugation the supernatant above and the cushion below is removed, leaving a concentrated sperm pellet in the conical ... It is sold under the trade name Visipaque; it is also sold as a density gradient medium under the name OptiPrep. Visipaque is ... 2008;70:704-714 https://web.archive.org/web/20160825063320/http://www.cosmobiousa.com/axis-shield-density-gradient-optiprep. ...
"Purification of plant mitochondria by isopycnic centrifugation in density gradients of Percoll". Arch Biochem Biophys. (1982) ...
It is also a density gradient medium for the centrifugation of biological particles.[citation needed] Historically metrizamide ...
Isolate the mRNA-ribosome complexes using sucrose gradient density centrifugation or specialized chromatography columns. Phenol ... These sites of slow or paused translation are demonstrated by an increase in ribosome density and these pauses can link ...
Hadjiolov AA, Venkov PV, Tsanev RG (November 1966). "Ribonucleic acids fractionation by density-gradient centrifugation and by ...
Density gradients. In this technique, a tube is filled with layers of liquids of different densities and semen is placed on the ... Then, The tube goes through a centrifugation to filter cell debris and non motile cells. After the centrifugation, healthy ... There are different techniques to perform the capacitation step: simple washing, migration (swim-up), density gradients, and ... "Density gradient capacitation is the most suitable method to improve fertilization and to reduce DNA fragmentation positive ...
"Separation and concentration of living dinoflagellate resting cysts from marine sediments via density-gradient centrifugation ... Another method, rarely used, uses a sucrose gradient. Recent times have brought about the possibility to get molecular ... calibration as annual density proxy and first evidence of pseudo-cryptic speciation". Journal of Quaternary Science. 27 (7): ...
Ycf4 co-fractionates with a protein complex larger than PSI upon sucrose density gradient centrifugation of solubilised ...
Density gradient centrifugation (in a continuous or discontinuous gradient) can concentrate semen samples with low ... comparison with density gradient centrifugation". Human Reproduction. 23 (12): 2646-2651. doi:10.1093/humrep/den330. ISSN 0268- ... which in combination with density gradient centrifugation in single sperm preparation protocols results in spermatozoa with ... "Selection of sperm based on combined density gradient and Zeta method may improve ICSI outcome". Human Reproduction. 24 (10): ...
... by density gradient centrifugation. The tube contains an insert that prevents the mixing of the density gradient medium and ... These immunorosettes pellet when centrifuged over a density gradient medium, leaving untouched target cells at the plasma: ... density gradient medium interface. SepMate is a tube that allows for the isolation of peripheral blood mononuclear cells (PBMC ...
... for density gradient centrifugation and for separating mixtures. Common applications of heavy liquids include: Density gradient ... A heavy liquid is a solution or liquid chemical substance with a high density and a relatively low viscosity. Heavy liquids are ... centrifugation Separating mixtures and sink/swim analysis Flotation process Determination of density The classical heavy ... With this relatively new heavy liquid densities up to 3.1 g·cm−3 can be adjusted . Adding parts of pulverulent Tungsten carbide ...
... or other substances on the basis of their density. Isopycnic centrifugation refers to a method wherein a density gradient is ... After this gradient is formed particles move within the gradient to the position having a density matching their own (this is ... Unless there is a flux of mass into or out of a control volume, a process which occurs at a constant density also occurs at a ... It may also be applied to other situations where a continuous medium has smoothly varying density, such as in the case of an ...
Sperm may be washed by density gradient centrifugation or by a "direct swim-up" technique that does not involve centrifugation ... In normal semen samples, centrifugation causes no more DNA damage to spermatozoa than a direct swim-up technique. Washed sperm ...
Such an "equilibrium" centrifugation can allow extensive purification of a given particle. Sucrose gradient centrifugation - a ... Centrifugation is a process that uses centrifugal force to separate mixtures of particles of varying masses or densities ... However, when the proteins are moving through a sucrose gradient, they encounter liquid of increasing density and viscosity. A ... Samples separated by these gradients are referred to as "rate zonal" centrifugations. After separating the protein/particles, ...
These studies utilized the method of density gradient centrifugation that was developed for the test of the semiconservative ...
These nuclei can then be removed using cytochalasin B to disrupt the cytoskeleton and centrifugation in a density gradient to ...
... and density gradient centrifugation. It has significant advantages when compared to zinc chloride solution or the toxic ... density 3.1 g/cm3), SPT is widely used as to produce "heavy liquid" for gravity separation (sink /float analysis) ...
Density gradient centrifugation is also another method for selecting flakes based on lateral size, where a density gradient is ... In the case of sorting MXenes, a sucrose and water density gradient can be used from 10 to 66 w/v %. Using density gradients ... differential centrifugation, and density gradient centrifugation procedures. Post processing methods rely heavily on the as- ... Differential centrifugation, also known as cascading centrifugation, can be used to select flakes based on lateral size by ...
... "density-gradient") centrifugation. It is isostructural with potassium salt. Coordination sphere of one of two types of Cs+ site ...
... and allowed them to be isolated as sealed structures by the combination of differential and density gradient centrifugation. He ... shock he subsequently showed that intact synaptic vesicles of high purity can be isolated by density gradient centrifugation ... an electron-microscopic study of cell fragments derived by homogenization and centrifugation". J Anat. 96 (Pt. 1): 79-88. PMC ...
... and TEM-8 in colorectal carcinoma patients using OncoQuick density gradient centrifugation system". J. Surg. Res. 155 (2): 183- ... 2007). "Relationship between vascular invasion and microvessel density and micrometastasis". World J. Gastroenterol. 13 (46): ...
... coli through sucrose-density-gradient centrifugation. Cell-free synthetic pathway biotransformation biosystems are proposed as ... Zhu Z, Kin Tam T, Sun F, You C, Percival Zhang YH (2014). "A high-energy-density sugar biobattery based on a synthetic ... reticulocytes have been lysed in a solution of MgCl2 and had the extract filtered away from the membranes by centrifugation. ...
... anticoagulated blood sample that contains most of the white blood cells and platelets following density gradient centrifugation ... Some refer to the buffy coat when separating cells by Ficoll density gradients. This usage of the expression buffy coat is ... After centrifugation, one can distinguish a layer of clear fluid (the plasma), a layer of red fluid containing most of the red ...
Blevins, B.A., de la Rey, M. and Loskutoff, N.M. (2008) Effect of density gradient centrifugation with trypsin on the ...
... to select sperm from semen by density gradient centrifugation, for use in techniques such as in vitro fertilization or ... It is used for the isolation of cells, organelles, and/or viruses by density centrifugation. Percoll consists of colloidal ... Pertoft, H.; Laurent, T.C.; Laas, T.; Kagedal, L. (1978). "Density gradients prepared from colloidal silica particles coated by ... Percoll is well suited for density gradient experiments because it possesses a low viscosity compared to alternatives, a low ...
Different types of EVs may be separated based on density[8]:Table 1 (by gradient differential centrifugation), size, or surface ... can be isolated by precise high-speed centrifugation in the density gradient. Using osmotic shock, it is possible temporarily ... Applying ionophores like valinomycin can create electrochemical gradients comparable to the gradients inside living cells. ... Big fragments of the crushed cells can be discarded by low-speed centrifugation and later the fraction of the known origin ( ...
The aim of the study was to compare the new density gradient centrifugation system OncoQuick with the standard density gradient ... centrifugation system Ficoll for improved tumor cell enrichment in... ... Density gradient centrifugation with OncoQuick results in higher relative tumor cell enrichment than Ficoll density gradient ... The aim of the study was to compare the new density gradient centrifugation system OncoQuick with the standard density gradient ...
Alternatively, density gradient centrifugation may be performed using multiple layers of the different gradient densities. This ... on the accuracy of the density gradients, i.e., the density gradients must be prepared such that their densities are accurate ... Density gradient centrifugation is a method of separating cells based on the different densities of cell types in a mixture. ... Thus, density gradient centrifugation is most often carried out through repetitive steps based on a series of different density ...
DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation, Protocol for Isolation of ... Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation, ... Isolation of Human Monocytes by Double Gradient Centrifugation and Their Differentiation to Macrophages in Teflon-coated Cell ... Preparation of Synaptic Plasma Membrane and Postsynaptic Density Proteins Using a Discontinuous Sucrose Gradient, Human ...
Density-Gradient Separation of Organic and Inorganic Particles by Centrifugation Message Subject. (Your Name) has forwarded a ... Aqueous suspensions of particulate organic and inorganic material were centrifuged in a 2-bromo-ethanol density gradient. The ... degree of separation with this new technique is superior to that achieved with sucrose density gradients. ...
Specialized tube for rapid PBMC isolation by density gradient centrifugation. Manufactured under cGMP and can be used for IVD ... into your density gradient centrifugation step. SepMate™ tubes contain an insert that creates a barrier between the density ... Recovery of MNCs from fresh and 48-hour post blood draw enriched by density gradient centrifugation with SepMate™ (purple) or ... How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™) ...
Localization of the Proton Pump of Corn Coleoptile Microsomal Membranes by Density Gradient Centrifugation. Suzanne Mandala, ... Localization of the Proton Pump of Corn Coleoptile Microsomal Membranes by Density Gradient Centrifugation ... Localization of the Proton Pump of Corn Coleoptile Microsomal Membranes by Density Gradient Centrifugation ... Localization of the Proton Pump of Corn Coleoptile Microsomal Membranes by Density Gradient Centrifugation ...
Similar isolations may be accomplished with density gradients in a swinging-bucket rotor. On isopycnic-density-gradient ... Isolation and partial characterization of high-density lipoprotein HDL1 from rat plasma by gradient centrifugation L T Lusk; L ... Isolation and partial characterization of high-density lipoprotein HDL1 from rat plasma by gradient centrifugation. Biochem J 1 ... More that 98% of the apoprotein of the lighter component was B protein, and hence this particle is LD (low-density) lipoprotein ...
What is density-gradient centrifugation? Meaning of density-gradient centrifugation medical term. What does density-gradient ... Looking for online definition of density-gradient centrifugation in the Medical Dictionary? density-gradient centrifugation ... density-gradient centrifugation. density-gradient centrifugation. the separation of the components of an homogenate by a ... and Percoll density-gradient centrifugation [13, 25].. Percoll density-gradient centrifugation method could be used to assess ...
See how to layer blood over the density medium and learn two different... ... This protocol describes how to isolate mononuclear cells from whole blood using density gradient centrifugation. ...
Zone-spreading of a major component by non-ideal sedimentation in density-gradient centrifugation can cause zone spreading of a ... but in a much wider zone after centrifugation with a large amount of a second virus. ... A small amount of southern bean mosaic virus was contained in a narrow zone after density-gradient centrifugation, ... A small amount of southern bean mosaic virus was contained in a narrow zone after density-gradient centrifugation, but in a ...
... subjecting the blood of the mother body to first density-gradient centrifugation to collect a cell fraction containing the ... subjecting the treated cell fraction containing the nucleated red blood cells to second density-gradient centrifugation to ... thereby avoiding the overlapping of the density of the nucleated red blood cells and the density of white blood cells each ... treating the cell fraction containing the nucleated red blood cells in such a manner that the density of the nucleated red ...
The main conclusions derived from this study were that 1) isolation of sperm from the gradient pellet by DGC must be performed ... The results showed that while there was a significant reduction in the SDF levels in sperm isolated from the gradient ... This was a prospective multicenter study aiming at comparing the efficiency of sperm selection by density gradient ... centrifugation (DGC) in reducing sperm DNA fragmentation (SDF) in different ART centers. The study was designed using 290 semen ...
... multi-step density gradient centrifugation techniques, as eosinophils have the highest density of all leukocytes, overlapping ... Single-site, five-year experience with human eosinophil isolation by density gradient centrifugation and CD16 immunomagnetic ... Therefore, the described method for human eosinophil isolation from whole blood using density gradient centrifugation, RBC ... year period with 350 consecutive human eosinophil isolation preparations from whole blood using density gradient centrifugation ...
Centrifugation, Density Gradient*Centrifugation, Density Gradient. *Centrifugations, Density Gradient. *Density Gradient ... Density Gradient" by people in this website by year, and whether "Centrifugation, Density Gradient" was a major or minor topic ... "Centrifugation, Density Gradient" is a descriptor in the National Library of Medicines controlled vocabulary thesaurus, MeSH ( ... Below are the most recent publications written about "Centrifugation, Density Gradient" by people in Profiles. ...
... depending on the density gradient media.. Please select your region. This is important to be able to checkout with correct ... During centrifugation, Leukocytes, lymphocytes and PBMCs are separated from unwanted erythrocytes and granulocytes, ...
Separation using density gradient centrifugation. Related Stories. *Biomarkers in the blood can predict severe outcomes in ... The diluted blood is then slowly added to density gradient media and centrifugation is performed for around 20 minutes. The ... New sterile conical tubes with same volume as density centrifugation should be used to contain the PBMCs and as much of the ... This should be done without the brakes, as de-acceleration can disrupt the density gradient formation. The centrifuge should ...
Density Gradient Centrifugation. 2 × 107 uninfected or infected HL-60 cells were washed with ice-cold 1x PBS twice followed by ... were homogenized and the post-nuclear supernatants were separated by density gradient centrifugation. Successive one-ml ... Lysates of HL-60 cells, host cell-free A. phagocytophilum organisms, or gradient centrifugation fractions were analyzed by SDS- ... phagocytophilum infected HL-60 cells were fractionated by a continuous density gradient fractionation method that keeps the ApV ...
... followed by density gradient centrifugation using Larex (Stractan) or other gradient medium to remove other non-parenchymal ... Isolation of human HSC (fat-storing cells) was also reported in other studies using density gradient centrifugation method [31 ... Isolation of human HSCs using density gradient centrifugation. Friedman [23] first successfully isolated, cultured, and ... Isolation and culture of hepatic lipocytes, Kupffer cells, and sinusoidal endothelial cells by density gradient centrifugation ...
Sucrose density gradient centrifugation.. This was done according to a method described previously (6). In brief, INS cell ... INS cell cytosolic fraction was subjected to sucrose (5-20%) density gradient centrifugation (see research design and methods ... To this end, using Triton X-114 phase partition, co-immunoprecipitation, and sucrose density gradient centrifugation approaches ... Western blot analysis of individual fractions isolated from β-cell lysates subjected to sucrose density gradient centrifugation ...
Density Gradient CentrifugationEdit. Density gradient centrifugation is considered one of the more efficient methods of ... Density gradient centrifugation can be used both as a separation technique and as a method of measuring the densities of ... which in fact have little to no density gradient.[7]. Differential CentrifugationEdit. Differential Centrifugation is a type of ... They used density gradient centrifugation to determine which isotope or isotopes of nitrogen were present in the DNA after ...
Sucrose density gradient centrifugation of TX-100-insoluble fraction. Continuous sucrose density gradients (5-60% or 5-40%) in ... 1994) by sucrose density gradient centrifugation. Both fractions, in accordance with their detergent insolubility, densities ... The sucrose density gradient fractions from centrifugation of the pellet obtained by procedure 1 had very little actin or ... aDensity of spot on TLC plate as a percentage of total densities of all spots. bRatio of densities of cholesterol (chol), GalC ...
... obtained from Neurospora crassa grown under different conditions was investigated by density-gradient centrifugation on Percoll ... density 1·059; to 1·061; g ml-1) was also present in the stationary phase. When N. crassa was grown with acetate as the carbon ... The glyoxysomes were found to be less dense than the mitochondria and their density decreased from 1·076; to 1·055; g ml-1 as ... The buoyant density of isolated mitochondria and glyoxysomes ... conditions was investigated by density-gradient centrifugation ...
Apoptosis , Centrifugation , Centrifugation, Density Gradient , Chromatin , DNA Fragmentation , DNA , Methods , Product ... Centrifugation / Centrifugation, Density Gradient / Apoptosis / Product Packaging / DNA Fragmentation Clinical aspect: ... and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing ... isolation of sperm with high DNA integrity and stable chromatin packaging by a combination of density-gradient centrifugation ...
Sucrose density gradient centrifugation. The cells were plated on 10-cm plates and transfected with 9 μg of total DNA per well ... A widely used approach in this category employs density gradient fractionation using either non-ionic detergent-based or ... Lubrol and separated into DRM and non-DRM fractions by sucrose gradient density fractionation at 180,000 × g for 18 h at 4°C. ... Lubrol and separated into DRM and non-DRM fractions by sucrose gradient density fractionation at 180,000 × g for 18 h at 4°C. ...
Keywords: Aneuploidy; Density gradient centrifugation; DNA fragmentation; Sperm; Swim-up Introduction. The percentage of sperm ... SU, swim-up; DGC, density gradient centrifugation.. a)Sperm cells with a small halo or no halo, or that were degraded, when ... Swim-up and density gradient centrifugation (DGC) are commonly used sperm preparation methods for assisted conception [5]. ... The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount ...
Density gradient centrifugation. Mitochondria were solubilized at 2 mg protein/ml in digitonin buffer for 20 min on ice and ... Furthermore, glycerol density gradient centrifugation of digitonin-solubilized mitochondria showed that Ups1p-Flag and Ups3p- ... Isolated mitochondria were solubilized in 1% digitonin and subjected to 20-40% glycerol density gradient centrifugation. In ... E) Mitochondria were solubilized with digitonin and subjected to glycerol density gradient centrifugation. Fractions were ...
OptiPrep density gradient centrifugation. The procedure was performed as previously described (Lobb et al., 2015) by using the ... 3 c). Together with the data on vesicle size, their buoyancies in density gradient centrifugation and their neutral ... 2 g). Subfractionation of EEVs by density gradient centrifugation and subsequent analyses by SDS-PAGE, Coomassie blue staining ... a buoyancy of 1.16-1.23 g/ml in density gradient centrifugation (Théry et al., 2006; Lobb et al., 2015), and a protein ...
Preparation of Enriched Plasma Membranes by Density Gradient Centrifugation-. A total of 5 × 108 cells were homogenized in 10 ... Percoll was removed by centrifugation of the fractions in 1-ml 1PC tubes at 900,000 × g in Sorvall RC M150 GX using the S150AT ... The gradient was fractionated from the top by the displacement method. In order to select fractions containing enriched plasma ... Homogenization and centrifugation were repeated. The post-nuclear supernatant was centrifuged at 50,000 × g for 30 min. The ...
Density gradient centrifugation. Lec-1 cells stably expressing GM or transiently expressing CD4 or sucrase-isomaltase were ... Our sucrose-density gradient analysis showed that an ERGIC-53 construct unable to form intermolecular disulfide bridges is ... It is difficult to correctly predict the size of the GM complexes from density gradients because the marker proteins are all ... Density gradient analysis may, therefore, underestimate the actual size of the GM complexes. Nevertheless, these experiments ...
Density Gradient Centrifugation by AUTHOR, UNKNOWN * PDF * £51.99 * Vitamin E Chemistry and Nutritional Benefits ...
  • X and Y spermatozoa can be separated using sorting methods such as flow cytometry, albumin sedimentation, and Percoll density-gradient centrifugation [13, 25]. (thefreedictionary.com)
  • Percoll density-gradient centrifugation method could be used to assess economic value and led to greater ease of use [27]. (thefreedictionary.com)
  • Sexing using Percoll density-gradient centrifugation method for 5 and 7 min was applied in this research. (thefreedictionary.com)
  • The buoyant density of isolated mitochondria and glyoxysomes obtained from Neurospora crassa grown under different conditions was investigated by density-gradient centrifugation on Percoll. (microbiologyresearch.org)
  • A simple, quick and reproducible method consisting of density-gradient centrifugation of homogenized infected mouse brain tissue on Percoll is described for the isolation and purification of cysts of Isospora (Toxoplasma) gondii. (nih.gov)
  • A 100% recovery of cysts, with 74.2% in a single fraction with a specific gravity of 1.056, was obtained by overlaying homogenates of infected mouse brains on a pre-formed Percoll gradient and centrifugation at low g forces. (nih.gov)
  • A rapid procedure for the isolation of lysosomes from kidney cortex by Percoll density gradient centrifugation. (bvsalud.org)
  • Highly pure lysosomes were isolated from buffalo ( Bubalus bubalis) kidney cortex by a procedure involving differential and isopycnic Percoll density gradient centrifugations . (bvsalud.org)
  • The highest purity of the isolated anammox consortium is 99% obtained by the method of percoll density gradient centrifugation [ 8 ]. (omicsonline.org)
  • Separation of particles according to density by employing a gradient of varying densities. (jove.com)
  • The degree of separation with this new technique is superior to that achieved with sucrose density gradients. (sciencemag.org)
  • the separation of the components of an homogenate by a process of CENTRIFUGATION where each component is moved to a level equivalent to its density in the solute gradient used. (thefreedictionary.com)
  • Centrifugal Separations in Molecular and Cell Biology focuses on the application of modern centrifugation technology in molecular and cell biology, including the separation and fractionation of biological particles by centrifugation on the preparative and analytical scales. (elsevier.com)
  • Topics include separation by isopycnic banding, large-scale preparative procedures, and density-gradient solutes. (elsevier.com)
  • Importantly, preparative ultracentrifugation reaches the g-force necessary for timely separation of biological macromolecule samples to their isopycnic point in density gradients. (pharmpro.com)
  • The three main methods of separation are differential pelleting, rate‐zonal centrifugation and isopycnic centrifugation. (els.net)
  • The choice of centrifugation method depends on the nature of the particles and often more than one separation technique is required. (els.net)
  • Many centrifugation purification procedures involve sequential centrifugation steps, often a size separation followed by purification using isopycnic gradient centrifugation. (els.net)
  • The figure describes the separation of three types of particle: red, green and magenta (density increasing in that order). (els.net)
  • The technique involves partial separation of organelles by density gradient centrifugation followed by the analysis of protein distributions in the gradient by ICAT and MS. Multivariate data analysis techniques are then used to group proteins according to their distributions and hence localizations. (mcponline.org)
  • Zone-spreading of a major component by non-ideal sedimentation in density-gradient centrifugation can cause zone spreading of a minor component that the major component overlaps. (unl.edu)
  • Using siRNAs originally designed and an alkaline sucrose density gradient sedimentation technique, we verified the involvement of several polymerases in ultraviolet (UV) light-induced TLS in HeLa cells. (pubmedcentralcanada.ca)
  • Discussions focus on the basic concepts of sedimentation theory, centrifugation methods, designing centrifugation experiments, care of centrifuges and rotors, and statistical estimation of molecular parameters. (elsevier.com)
  • Many methods have been developed for separating human spermatozoa from seminal plasma, including the swim-up (SUP) method, self-migration sedimentation, glass wool filtration, magnetic-activated cell sorting (MACS), and the use of a density gradient medium [ 4 ]. (ecerm.org)
  • The results of subcellular fractionation and sedimentation in density gradients of extracts prepared from postdivisional sporangia confirmed that pro-ς E was chiefly present in the membrane fraction and that ς E was predominantly cytoplasmic, findings that suggest that the pro-amino acid sequence is responsible for the sequestration of pro-ς E to the membrane. (asm.org)
  • Rate‐zonal or sedimentation velocity density gradient centrifugation. (els.net)
  • Lymphocyte thymidine uptake in leucocytes prepared by NH 4 Cl induced RBC lysis was compared with that of lymphocytes obtained following Ficoll-Hypaque centrifugation using polynomial regression dose response curves which adjusted for inherent subject variability. (elsevier.com)
  • During centrifugation, Leukocytes, lymphocytes and PBMCs are separated from unwanted erythrocytes and granulocytes, depending on the density gradient media. (pluriselect.com)
  • Isolation of Tumor-Infiltrating Lymphocytes by Ficoll-Paque Density Gradient Centrifugation. (nih.gov)
  • following centrifugation, lymphocytes are collected from the plasma-Ficoll interface. (drugs.com)
  • To this end, using Triton X-114 phase partition, co-immunoprecipitation, and sucrose density gradient centrifugation approaches, we report coexistence of GDI with Rac1 in insulin-secreting β-cells (INS cells). (diabetesjournals.org)
  • In contrast, Y348A receptors did not undergo "endocytosis" into the light vesicle fraction in sucrose density gradient centrifugation assays, as did the wild-type receptor. (aspetjournals.org)
  • The aim of the study was to compare the new density gradient centrifugation system OncoQuick with the standard density gradient centrifugation system Ficoll for improved tumor cell enrichment in blood of tumor patients. (springer.com)
  • isolated by using standard density gradient centrifugation. (cdc.gov)
  • Using rate zonal ultracentrifugation, we isolated two lipoproteins in almost equal proportions from this density range. (portlandpress.com)
  • On isopycnic-density-gradient ultracentrifugation one component banded at rho = 1.031 g/ml and the other at rho = 1.054 g/ml. (portlandpress.com)
  • HDL was subfractionated by density gradient ultracentrifugation, and LpA-I was analyzed by electroimmunodiffusion. (nih.gov)
  • Additionally, exosomes have been shown to be part of intercellular communication functions, with implications toward both anti-tumor and pro-tumor activity.6 Previous work has provided insight to the isolation of exosomes using density gradient ultracentrifugation,2,7 although there is an effort to gain more concrete confirmation of the size and concentration after purification. (pharmpro.com)
  • 1999) Purification of recombinant adeno‐associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system. (els.net)
  • Owing to the association of S protein with the thrombin-antithrombin III (TAT) complex, the STAT complex assembled in vitro exhibited a higher Mr than the TAT complex as judged by polyacrylamide-gradient-gel electrophoresis in the absence of SDS. (biomedsearch.com)
  • Most of these techniques involve carrying out numerous operations (e.g., pipetting, centrifugations, electrophoresis) on a large number of samples. (google.ca)
  • From 10 ml blood, OncoQuick density gradient centrifugation separated a cell fraction which consisted of a mean cell number of 9.5×l0 4 mononuclear cells compared to 1.8×l0 7 cells by Ficoll. (springer.com)
  • SepMate™ tubes contain an insert that creates a barrier between the density gradient medium and blood, thus eliminating the need for careful blood layering and allowing mononuclear cells to be easily harvested with a simple pour. (stemcell.com)
  • The MACSprep™ PBMC Isolation Kit has been developed for the fast isolation of human peripheral blood mononuclear cells (PBMCs) from 1-8 mL of freshly drawn anticoagulated whole blood without density gradient centrifugation. (miltenyibiotec.com)
  • The book also ponders on the practical aspects of rate-zonal centrifugation, including gradient materials, density and viscosity of glycerol solutions, and resolution and gradient shape. (elsevier.com)
  • The publication examines fractionations in zonal rotors and the quantitative aspects of rate-zonal centrifugation. (elsevier.com)
  • Particles can be separated on the basis of their size using differential pelleting or rate‐zonal centrifugation. (els.net)
  • The text then reviews isopycnic centrifugation in ionic media and analytical centrifugation. (elsevier.com)
  • The first two methods separate particles primarily on the basis of size while isopycnic centrifugation separates particles on the basis of their density. (els.net)
  • For example, membrane fractionation often involves first making an enriched fraction from a cell homogenate by differential pelleting followed by isopycnic centrifugation to obtain purified fractions. (els.net)
  • Alternatively, particles can be separated on the basis of their density using isopycnic centrifugation. (els.net)
  • Early attempts at purifying nucleic acids relied on density-gradient centrifugation , an expensive process that requires multiple steps and dangerous chemicals. (thefreedictionary.com)
  • Centrifugation is the use of the centrifugal forces generated in a spinning rotor to separate biological particles, such as cells, viruses, sub‐cellular organelles, macromolecules (principally proteins and nucleic acids) and macromolecular complexes (such as ribonucleoproteins and lipoproteins). (els.net)
  • These samples would be pooled and processed by using available technology to isolate virus particles en masse, recover viral nucleic acids, produce amplified shotgun libraries, carry out shotgun sequencing of the mixture of viral genomes, and reconstruct these genomes in silico with the techniques originally developed to sequence the entire human genome from random fragments. (cdc.gov)
  • New sterile conical tubes with same volume as density centrifugation should be used to contain the PBMCs and as much of the plasma as possible should be removed. (news-medical.net)
  • PBMCs were plated at a density of 2.5 × 105 cells/well. (cdc.gov)
  • Centrifugation is a technique which involves the application of centrifugal force to separate particles from a solution according to their size, shape, density, viscosity of the medium and rotor speed. (wikipedia.org)
  • The particles' settling velocity in centrifugation is a function of their size and shape, centrifugal acceleration, the volume fraction of solids present, the density difference between the particle and the liquid, and the viscosity . (wikipedia.org)
  • The selection first covers the principles and practices of centrifugation and the bases of centrifugal separations. (elsevier.com)
  • Preparative Centrifugation - a Practical Approach, pp. 77-142. (els.net)
  • Preparative Centrifugation: A Practical Approach (1992) Edited D Rickwood , Oxford: Oxford University Press. (els.net)
  • One solution to this problem is the use of analytical rather than preparative centrifugation. (mcponline.org)
  • Analytical centrifugation is an established method for assigning individual proteins to subcellular compartments that have eluded purification and refers to the analysis of protein distributions in density gradients, as opposed to preparative centrifugation, which is the analysis of single organelle-enriched fractions ( 3 , 4 ). (mcponline.org)
  • Evaluation of OncoQuick and Ficoll density gradient centrifugation was performed by flow-cytometry and immunocytochemistry using 10 ml unspiked and tumor cell-spiked blood samples of tumor-free probands. (springer.com)
  • Density gradient centrifugation of tumor cell-spiked blood samples with OncoQuick and Ficoll led to similar tumor cell recovery rates, between 70% and 90% for both methods. (springer.com)
  • Consequently, tumor cell density on each cytospin was higher after OncoQuick preparation compared to Ficoll. (springer.com)
  • Density gradient centrifugation with OncoQuick results in higher relative tumor cell enrichment than Ficoll density gradient centrifugation. (springer.com)
  • See DIFFERENTIAL CENTRIFUGATION , ULTRACENTRIFUGE , EQUILIBRIUM CENTRIFUGATION . (thefreedictionary.com)
  • Flow chart of a typical differential centrifugation of a cell or tissue homogenate. (els.net)
  • Differential centrifugation of a homogenate: (a) before centrifugation and (b) after centrifugation. (els.net)
  • Gertler R., Rosenberg R., Fuehrer K., Dahm M., Nekarda H., Siewert J.R. (2003) Detection of Circulating Tumor Cells in Blood Using an Optimized Density Gradient Centrifugation. (springer.com)
  • En route to their target organs migratory cells have to overcome various architectural hindrances, including interstitial matrix structures, basement membranes, and intercellular junctions. (rupress.org)
  • The resulting suspension of cells is sedimented through a sucrose density gradient. (biomedsearch.com)
  • Depending on the centrifugation force used, particles from cells to those as small as macromolecules can be separated. (els.net)
  • To investigate the effect of thyroid dysfunction on high-density lipoprotein (HDL) metabolism, we measured HDL subfractions, apolipoprotein A-I containing particles (LpA-I and LpA-I:A-II), and the activities of enzymes involved in the remodeling and metabolism of HDL [namely hepatic lipase (HL), lipoprotein lipase, and cholesteryl ester transfer protein (CETP)] in 18 hyperthyroid and 17 hypothyroid patients before and after treatment. (nih.gov)
  • We have developed a proteomics technique for the localization of integral membrane proteins to compartments of the endomembrane system in Arabidopsis thaliana that is based upon analytical centrifugation and hence is not dependent on the production of pure organelles. (mcponline.org)
  • Barry, "Use of potassium tartrate for equilibrium density-gradient centrifugation of animal viruses," Nature, vol. (thefreedictionary.com)
  • At equilibrium each particle settles in the gradient at a point equal to its density. (childrensmercy.org)
  • Wetter, 1960), ( b ) further fractionation by rate or equilibrium density gradient centrifugation as developed by Brakke (1960). (microbiologyresearch.org)
  • More that 98% of the apoprotein of the lighter component was B protein, and hence this particle is LD (low-density) lipoprotein. (portlandpress.com)
  • There is a correlation between the size and density of a particle and the rate that the particle separates from a heterogeneous mixture, when the only force applied is that of gravity. (wikipedia.org)
  • Each particle sediments to the position at which the gradient density is equal to its own. (bioportfolio.com)
  • Centrifugation, Density Gradient" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings) . (childrensmercy.org)
  • These methods initially involved intricate, multi-step density gradient centrifugation techniques, as eosinophils have the highest density of all leukocytes, overlapping in this regard only with neutrophils [ 11 ]. (biomedcentral.com)
  • Two different types of low-density detergent-insoluble glycosphingolipid-enriched membrane domain (DIG) fractions were isolated from myelin by extraction with Triton X-100 (TX-100) in 50 mM sodium phosphate buffer at room temperature (20°C) (procedure 1), in contrast to a single low-density fraction obtained by extraction with TX-100 in Tris buffer containing 150 mM NaCl and 5 mM EDTA at 4°C (procedure 2). (wiley.com)
  • The single DIG fraction obtained by procedure 2 gave a single opaque band at a similar sucrose density to B1. (wiley.com)
  • Both B1 and B2 had characteristics of lipid rafts, i.e. high galactosylceramide and cholesterol content and enrichment in GPI-linked 120-kDa neural cell adhesion molecule (NCAM)120, as found by others for the single low-density DIG fraction obtained by procedure 2. (wiley.com)
  • The single low-density DIG fraction obtained by procedure 2 contained only low amounts of actin and tubulin. (wiley.com)
  • Buoyant density gradient centrifugation in a continuous gradient. (els.net)
  • A technique used to separate particles according to their densities in a continuous density gradient. (bioportfolio.com)
  • The method has flexibility to change volumes for each gradient, as well as the number of tubes prepared. (pharmpro.com)
  • HIV-1 p24-Gag and Nef contained the highest epitope density and were also the most frequently recognized HIV-1 proteins. (asm.org)
  • Proteins from each organelle co-fractionate and therefore exhibit similar distributions in the gradient. (mcponline.org)
  • Protein distributions can be determined through a series of pair-wise comparisons of gradient fractions, using cleavable ICAT to enable relative quantitation of protein levels by MS. The localization of novel proteins is determined using multivariate data analysis techniques to match their distributions to those of proteins that are known to reside in specific organelles. (mcponline.org)
  • This is particularly problematic in the case of the endomembrane system, owing to the similar densities of its component organelles and the continuous cycling of membranes and proteins between these compartments. (mcponline.org)
  • Centrifugation of protein solution, for example, allows elimination of impurities into the supernatant. (wikipedia.org)
  • This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. (bvsalud.org)
  • The aim of this study was to compare the efficacy of swim-up and density gradient centrifugation (DGC) for reducing the amount of sperm with fragmented DNA, sex chromosome aneuploidy, and abnormal chromatin structure. (ecerm.org)
  • Swim-up and density gradient centrifugation (DGC) are commonly used sperm preparation methods for assisted conception [ 5 ]. (ecerm.org)
  • Ham's F-10 medium or sperm-washing medium (SWM) was used in combination with the density-gradient centrifugation/swim-up (DGC-SUP) or SUP methods for sperm preparation. (ecerm.org)
  • Apoptotic-like changes in equine spermatozoa separated by density-gradient centrifugation or after cryopreservation. (thefreedictionary.com)
  • Effect of density-gradient centrifugation on quality and recovery rate of equine spermatozoa. (thefreedictionary.com)
  • Specifically, the Biomek 4000 Workstation helps overcome human variables and provides a consistent, reproducible, high-throughput method for gradient setup, representing a breakthrough solution to scale-up problems. (pharmpro.com)
  • The Biomek 4000 Laboratory Automation Workstation helps overcome the human variable and provides a consistent, reproducible, high-throughput method for density gradient setup- an elegant solution to scale-up hurdles. (pharmpro.com)
  • Diagnostic value of longitudinal flow gradient for the presence of haemodynamically significant coronary artery disease. (bioportfolio.com)
  • This study aimed to investigate the diagnostic value of longitudinal MBF gradient for the presence of haemodynamically significant CAD. (bioportfolio.com)
  • This is a prospective, multi-center trial conducted at 1 Guangzhou and 2 Beijing centers designed to determine the diagnostic performance of virtual hepatic venous pressure gradient (vHVPG. (bioportfolio.com)
  • a) Sample is loaded in a narrow band on top of a continuous density gradient. (els.net)
  • These membranes were then fractionated according to their density by centrifuging at 350,000 × g for 3 h in a VTi65.1 rotor (Beckman Coulter), during which time an iodixanol gradient is generated. (mcponline.org)
  • Therefore, we have retrospectively reviewed our experience over a five-year period with 350 consecutive human eosinophil isolation preparations from whole blood using density gradient centrifugation, red blood cell (RBC) hypotonic lysis and immunomagnetic removal of contaminating neutrophils using CD16 antibody and herein describe key features of this methodologic approach. (biomedcentral.com)
  • Because of the small capacity of the swinging-bucket rotor generally employed only relatively small volumes of the partially purified preparations can be fractionated in the gradients. (microbiologyresearch.org)
  • In particular, the present invention relates to the use of a cell-trap centrifugation tube containing a specific density gradient solution adjusted to the specific density of a desired cell population to enrich for the desired cell from a cell source. (freepatentsonline.com)
  • In addition, the method can be further simplified by density-adjusted cell sorting which uses cell type-specific binding agents such as antibodies and lectins linked to carrier particles to impart a different density to the undesired populations in a more convenient manner. (freepatentsonline.com)
  • Centrifugation is the most common method used for uranium enrichment , relying on the slight mass difference between atoms of U238 and U235 in uranium hexafluoride gas. (wikipedia.org)
  • Centrifugation is also the most common method used for uranium enrichment, relying on the slight mass difference between atoms of U-238 and U-235 in uranium hexafluoride gas. (wikipedia.org)
  • Challenges to researchers working with exosomes include setting up density gradients by hand, because it is tedious, time-consuming and subject to user, lab, and method variability. (pharmpro.com)
  • Similar isolations may be accomplished with density gradients in a swinging-bucket rotor. (portlandpress.com)
  • The tube allows the desired cell population to be collected by decantation after centrifugation to minimize cell loss and maximize efficiency. (freepatentsonline.com)
  • A slow pipetting technique with liquid level sensing was used to minimize interface mixing of the gradients. (pharmpro.com)
  • Simplify peripheral blood mononuclear cell (PBMC) isolation by incorporating SepMate™ into your density gradient centrifugation step. (stemcell.com)
  • Eliminates the need for carefully layering blood over the density gradient medium (e.g. (stemcell.com)
  • The diluted blood is then slowly added to density gradient media and centrifugation is performed for around 20 minutes. (news-medical.net)
  • The longitudinal myocardial blood flow (MBF) gradient derived from positron emission tomography (PET) has been proposed as an emerging non-invasive index of haemodynamically significant coronary artery disease (CAD). (bioportfolio.com)
  • The two DIG fractions obtained by procedure 1 gave opaque bands, B1 and B2, at somewhat lower and higher sucrose density respectively than myelin itself. (wiley.com)
  • Fractions of 0.5 ml were harvested from the top of the gradient using an Auto Densi-flow collection device (Labconco Corporation, Kansas City, MO). (mcponline.org)
  • a rough guide to the 'g' force exerted at this speed is to multiply the centrifugation radius by a factor of 10, so a radius of 160 mm gives approximately 1600 x g. (wikipedia.org)
  • Leukemic blasts (mostly CD34 + ) from patients with acute myeloblastic leukemia (AML) expressed variable amounts of CXCR-4, which was functionally active, as demonstrated by a positive correlation between the SDF-1-induced transendothelial migration and the cell surface density of CXCR-4 (r = 0.97). (bloodjournal.org)
  • The amino acid sequence NPXY was shown to be an internalization signal for the low-density lipoprotein receptor ( 5 ), and related tyrosine-containing internalization signals are present in the intracellular domain of various other receptors that are internalized by a pathway involving clathrin-coated pits and vesicles ( 6 , 7 ). (aspetjournals.org)
  • The technique has a huge advantage over normal centrifugation, in that several components of different density can be separated in the same tube. (thefreedictionary.com)
  • Graham J , Ford T and Rickwood D (1994) The preparation of subcellular organelles from mouse liver in self‐generated gradients of iodixanol. (els.net)
  • Here we present a simple workflow using automation, centrifugation and dynamic light scattering (DLS), to purify and analyze exosome samples. (pharmpro.com)
  • Organelles are partially separated using centrifugation through self-generating density gradients. (mcponline.org)