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- Immunoelectrophoresis abstract "Immunoelectrophoresis is a general name for a number of biochemical methods for separation and characterization of proteins based on electrophoresis and reaction with antibodies. All variants of immunoelectrophoresis require immunoglobulins, also known as antibodies reacting with the proteins to be separated or characterized. The methods were developed and used extensively during the second half of the 20th century. In somewhat chronological order: Immunoelectrophoretic analysis (one-dimensional immunoelectrophoresis ad modum Grabar), crossed immunoelectrophoresis (two-dimensional quantitative immunoelectrophoresis ad modum Clarke and Freeman or ad modum Laurell), rocket-immunoelectrophoresis (one-dimensional quantitative immunoelectrophoresis ad modum Laurell), fused rocket immunoelectrophoresis ad modum Svendsen and Harboe, affinity immunoelectrophoresis ad modum Bøg-Hansen.Agarose as 1% gel slabs of about 1 mm thickness buffered at high pH (around 8.6) is traditionally preferred for the electrophoresis as well as the reaction with antibodies. The agarose was chosen as the gel matrix because it has large pores allowing free passage and separation of proteins, but provides an anchor for the immunoprecipitates of protein and specific antibodies. The high pH was chosen because antibodies are practically immobile at high pH. An electrophoresis equipment with a horizontal cooling plate was normally recommended for the electrophoresis.Immunoprecipitates may be seen in the wet agarose gel, but are stained with protein stains like Coomassie Brilliant Blue in the dried gel. In contrast to SDS-gel electrophoresis, the electrophoresis in agarose allows native conditions, preserving the native structure and activities of the proteins under investigation, therefore immunoelectrophoresis allows characterization of enzyme activities and ligand binding etc. in addition to electrophoretic separation.The immunoelectrophoretic analysis ad modum Grabar is the classical method of immunoelectrophoresis. Proteins are separated by electrophoresis, then antibodies are applied in a trough next to the separated proteins and immunoprecipitates are formed after a period of diffusion of the separated proteins and antibodies against each other. The introduction of the immunoelectrophoretic analysis gave a great boost to protein chemistry, some of the very first results were the resolution of proteins in biological fluids and biological extracts. Among the important observations made were the great number of different proteins in serum, the existence of several immunoglobulin classes and their electrophoretic heterogeneity.Crossed immunoelectrophoresis is also called two-dimensional quantitative immunoelectrophoresis ad modum Clarke and Freeman or ad modum Laurell. In this method the proteins are first separated during the first dimension electrophoresis, then instead of the diffusion towards the antibodies, the proteins are electrophoresed into an antibody-containing gel in the second dimension. Immunoprecipitation will take place during the second dimension electrophorsis andthe immunoprecipitates have a characteristic bell-shape, each precipitate representing one antigen, the position of the precipitate being dependent on the amount of protein as well as the amount of specific antibody in the gel, so relative quantification can be performed. The sensitivity and resolving power of crossed immunoelectrophoresis is than that of the classical immunoelectrophoretic analysis and there are multiple variations of the technique useful for various purposes. Crossed immunoelectrophoresis has been used for studies of proteins in biological fluids, particularly human serum, and biological extracts. Rocket immunoelectrophoresis is one-dimensional quantitative immunoelectrophoresis. The method has been used for quantitation of human serum proteins before automated methods became available.Fused rocket immunoelectrophoresis is a modification of one-dimensional quantitative immunoelectrophorsis used for detailed measurement of proteins in fractions from protein separation experiments.Affinity immunoelectrophoresis is based on changes in the electrophoretic pattern of proteins through specific interaction or complex formation with other macromolecules or ligands. Affinity immunoelectrophoresis has been used for estimation of binding constants, as for instance with lectins or for characterization of proteins with specific features like glycan content or ligand binding. Some variants of affinity immunoelectrophoresis are similar to affinity chromatography by use of immobilized ligands.The open structure of the immunoprecipitate in the agarose gel will allow additional binding of radioactively labeled antibodies to reveal specific proteins. This variation has been used for identification of allergens through reaction with IgE.Two factors determine that immunoelectrophoretic methods are not widely used. First they are rather work intensive and require some manual expertise. Second they require rather large amounts of polyclonal antibodies. Today gel electrophoresis followed by electroblotting is the preferred method for protein characterization because its ease of operation, its high sensitivity, and its low requirement for specific antibodies. In addition proteins are separated by gel electrophoresis on the basis of their apparent molecular weight, which is not accomplished by immunoelectrophoresis, but nevertheless immunoelectrophoretic methods are still useful when non-reducing conditions are needed.".
- Immunoelectrophoresis thumbnail CrossedimmunoelectrophoresisTCBH.jpg?width=300.
- Immunoelectrophoresis wikiPageExternalLink issuetoc.
- Immunoelectrophoresis wikiPageExternalLink immelec.htm.
- Immunoelectrophoresis wikiPageExternalLink 64.
- Immunoelectrophoresis wikiPageID "2428570".
- Immunoelectrophoresis wikiPageLength "7654".
- Immunoelectrophoresis wikiPageOutDegree "31".
- Immunoelectrophoresis wikiPageRevisionID "678597738".
- Immunoelectrophoresis wikiPageWikiLink Affinity_chromatography.
- Immunoelectrophoresis wikiPageWikiLink Affinity_electrophoresis.
- Immunoelectrophoresis wikiPageWikiLink Agarose.
- Immunoelectrophoresis wikiPageWikiLink Antibodies.
- Immunoelectrophoresis wikiPageWikiLink Antibody.
- Immunoelectrophoresis wikiPageWikiLink Binding_constant.
- Immunoelectrophoresis wikiPageWikiLink Category:Biochemistry_methods.
- Immunoelectrophoresis wikiPageWikiLink Category:Electrophoresis.
- Immunoelectrophoresis wikiPageWikiLink Category:Immunologic_tests.
- Immunoelectrophoresis wikiPageWikiLink Category:Laboratory_techniques.
- Immunoelectrophoresis wikiPageWikiLink Category:Molecular_biology.
- Immunoelectrophoresis wikiPageWikiLink Category:Protein_methods.
- Immunoelectrophoresis wikiPageWikiLink Complex_formation.
- Immunoelectrophoresis wikiPageWikiLink Coomassie_Brilliant_Blue.
- Immunoelectrophoresis wikiPageWikiLink Coordination_complex.
- Immunoelectrophoresis wikiPageWikiLink Electroblotting.
- Immunoelectrophoresis wikiPageWikiLink Electrophoresis.
- Immunoelectrophoresis wikiPageWikiLink Enzyme.
- Immunoelectrophoresis wikiPageWikiLink Gel_electrophoresis.
- Immunoelectrophoresis wikiPageWikiLink Glycan.
- Immunoelectrophoresis wikiPageWikiLink Immunoglobulins.
- Immunoelectrophoresis wikiPageWikiLink Lectin.
- Immunoelectrophoresis wikiPageWikiLink Ligand.
- Immunoelectrophoresis wikiPageWikiLink Ligands.
- Immunoelectrophoresis wikiPageWikiLink Macromolecule.
- Immunoelectrophoresis wikiPageWikiLink Macromolecules.
- Immunoelectrophoresis wikiPageWikiLink PH.
- Immunoelectrophoresis wikiPageWikiLink Protein.
- Immunoelectrophoresis wikiPageWikiLink File:CrossedimmunoelectrophoresisTCBH.jpg.
- Immunoelectrophoresis wikiPageWikiLink File:Plasmodium_glutamate_dehydrogenase_precipitation.jpg.
- Immunoelectrophoresis wikiPageWikiLinkText ""rocket immunoelectrophoresis"".
- Immunoelectrophoresis wikiPageWikiLinkText "Immunoelectrophoresis".
- Immunoelectrophoresis wikiPageWikiLinkText "affinity immunoelectrophoreis".
- Immunoelectrophoresis wikiPageWikiLinkText "immunochemical".
- Immunoelectrophoresis wikiPageWikiLinkText "immunoelectrophoresis".
- Immunoelectrophoresis hasPhotoCollection Immunoelectrophoresis.
- Immunoelectrophoresis wikiPageUsesTemplate Template:Immunologic_techniques_and_tests.
- Immunoelectrophoresis wikiPageUsesTemplate Template:MeshName.
- Immunoelectrophoresis wikiPageUsesTemplate Template:Reflist.
- Immunoelectrophoresis subject Category:Biochemistry_methods.
- Immunoelectrophoresis subject Category:Electrophoresis.
- Immunoelectrophoresis subject Category:Immunologic_tests.
- Immunoelectrophoresis subject Category:Laboratory_techniques.
- Immunoelectrophoresis subject Category:Molecular_biology.
- Immunoelectrophoresis subject Category:Protein_methods.
- Immunoelectrophoresis hypernym Name.
- Immunoelectrophoresis type Laboratory.
- Immunoelectrophoresis type Method.
- Immunoelectrophoresis type Process.
- Immunoelectrophoresis type Technique.
- Immunoelectrophoresis comment "Immunoelectrophoresis is a general name for a number of biochemical methods for separation and characterization of proteins based on electrophoresis and reaction with antibodies. All variants of immunoelectrophoresis require immunoglobulins, also known as antibodies reacting with the proteins to be separated or characterized. The methods were developed and used extensively during the second half of the 20th century.".
- Immunoelectrophoresis label "Immunoelectrophoresis".
- Immunoelectrophoresis sameAs Inmunoelectroforesis.
- Immunoelectrophoresis sameAs Immuno-électrophorèse.
- Immunoelectrophoresis sameAs Immunoelettroforesi.
- Immunoelectrophoresis sameAs m.07ccvg.
- Immunoelectrophoresis sameAs Q855786.
- Immunoelectrophoresis sameAs Q855786.
- Immunoelectrophoresis wasDerivedFrom Immunoelectrophoresis?oldid=678597738.
- Immunoelectrophoresis depiction CrossedimmunoelectrophoresisTCBH.jpg.
- Immunoelectrophoresis isPrimaryTopicOf Immunoelectrophoresis.