Technology
Affinity Chromatography
A chromatographic separation technique based on the ability of proteins to bind specific ligands reversibly.
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Technology
A chromatographic separation technique based on the ability of proteins to bind specific ligands reversibly.
Technology
An electrophoresis procedure, in which a sample is introduced into a capillary tube and the components are separated by the application of high voltage. Due to the large surface to volume ratio, cooling is more efficient, allowing use of higher voltages. As a result, separation occurs more quickly and sensitivity is increased. The technique allows for separation of proteins, nucleic acids, and carbohydrates.
Technology
A molecular genetic technique that combines chromatin immunoprecipitation (ChIP) with massively parallel DNA sequencing to map the binding sites of DNA-associated proteins in a sample of cells. First, crosslinked protein-DNA complexes are isolated using ChIP. Next, the crosslinks are broken, the proteins are removed and the purified DNA is modified with adaptor oligonucleotides to facilitate massively parallel DNA sequencing. Following sequencing, the DNA sequences that are obtained can be mapped to their genomic locations.
Technology
Gel electrophoresis is a method of separating large molecules (such as DNA fragments or proteins) from a mixture of similar molecules by electrophoretic filtration through a gelatinous matrix. Subject to an electric field, each molecule in the sample mixture migrates through the matrix at a different rate, depending mainly on electrical charge and size, resulting in spatial separation within the matrix of each molecular species in the sample mixture. Agarose and acrylamide in a cylindrical tube or slab gel are commonly used media for electrophoresis of proteins and nucleic acids.
Technology
A method for the detection of proteins or antigens on samples of cells that are grown in monolayers or in suspension and deposited on slides using antibodies or other biomolecules that bind the target and can be visualized through a chemical process.
Technology
An enhancement of immunoelectrophoresis in which a series of post-electrophoretic gel slabs are layered with cellulose-acetate gels saturated with specific antibodies. The resulting antigen-antibody complexes fixed on the second gel may then be stained, allowing sensitive and specific qualitative visual identification of paraproteins by electrophoretic position. It can be used to identify specific proteins in the blood or urine. It has greatest application in the identification and monitoring of monoclonal proteins that are produced in conditions like Waldenstom's macroglobulinaemia and multiple myeloma.
Technology
Analysis using microarray technology (e.g., cDNA arrays to see gene expression or protein microarrays to profile the pattern of proteins).
Technology
Analytical and separative procedures in which molecules, particularly proteins or nucleic acids, are separated by their different electrophoretic mobilities in a hydrated gel. The gel suppresses convective mixing of the fluid phase through which the electrophoresis takes place and contributes molecular sieving. For proteins, commonly carried out in the presence of the anionic detergent sodium dodecylsulphate (SDS). SDS denatures proteins so that noncovalently associating sub unit polypeptides migrate independently and by binding to the proteins confers a net negative charge roughly proportional to the chain weight.
Technology
Pulsed-field gel electrophoresis (PFGE) is a method used to separate proteins or high molecular weight linear DNA fragments that are greater than 50 kilobases (kb) in size. Fragments are separated by pulsing an electrical field across an agarose gel rather than applying the constant field used in conventional electrophoresis.
Technology
Sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis is a procedure for estimating molecular weights of polypeptides, and uses SDS, a detergent that dissociates and unfolds oligomeric proteins into its subunits. The SDS binds to the polypeptides to form complexes with fairly constant charge to mass ratios. The electrophoretic migration rate through a gel is therefore determined only by the size of the complexes.
Technology
A laboratory procedure in which proteins are first separated across a gel, usually polyacrylamide, according to their isoelectric point, and then separated in a perpendicular direction on the basis of their molecular weight.
Technology
A laboratory procedure for the detection or identification of proteins or peptides that have been separated by gel electrophoresis and transferred onto nitrocellulose or other type of paper or nylon membrane. The proteins are then detected by reaction with enzymatically labeled or radiolabeled antibody probes.