Raising monospecific antibodies by use of protein components prestained with Remazol Brilliant Blue and separated by disc electrophoresis on polyacrylamide gel

Autor:	A M Saoji, S S Kelkar, C Y Jad, Yemul Vl, P M Khare
Rok vydání:	1984
Předmět:	Radial immunodiffusion Chromatography medicine.diagnostic_test Chemistry Biochemistry (medical) Clinical Biochemistry Albumin Immunoelectrophoresis Gel electrophoresis of proteins Blood proteins Immunodiffusion Electrophoresis medicine Polyacrylamide gel electrophoresis
Zdroj:	Clinical Chemistry. 30:1252-1254
ISSN:	1530-8561 0009-9147
DOI:	10.1093/clinchem/30.7.1252
Popis:	We have reported (Clin Chem 29: 42-44, 1983) that prestaining with Remazol Brilliant Blue permits direct visualization of serum components on disc electrophoresis, and apparently purifies the proteins well. Here we have cut out the bands corresponding to the prestained albumin and transferrin after disc electrophoresis of normal human serum proteins, eluted some individual proteins into saline, and assessed their purity by immunoelectrophoresis and two-dimensional crossed immunoelectrophoresis against polyvalent antihuman serum. These two techniques indicated purity of these antigens. We inoculated rabbits with the eluates containing the pure antigens, and tested the resulting antibodies for monospecificity by immunoelectrophoresis, rocket electrophoresis, and single radial immunodiffusion. From the results we conclude that the antibodies raised against each component were monospecific, and that this is a simple, economical, rapid, and reliable method for obtaining a pure fraction of serum protein for use as an antigen.
Databáze:	OpenAIRE
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