Isoelectric focusing of human antibodies directed against a high molecular weight antigen. 1980

L Nye, and I M Roitt

A simple technique has been developed to assess the spectrotypes of antigen-specific human autoantibodies. Following isoelectric focusing in 3 M urea at 1200 V for 2 h, immunoglobulins were fixed in polyacrylamide gels by precipitation with sheep anti-human immunoglobulin in the presence of sodium sulphate. Specific anti-thyroglobulin antibody bands were labelled by reaction with [125I]thyroglobulin, an antigen with a molecular weight of 660,000 daltons which is too large to enter the gel. The advantages of this method include increased sensitivity and reproducibility, and the ability to produce sharper bands and less non-specific binding than other methods.

UI MeSH Term Description Entries
D007122 Immunoelectrophoresis A technique that combines protein electrophoresis and double immunodiffusion. In this procedure proteins are first separated by gel electrophoresis (usually agarose), then made visible by immunodiffusion of specific antibodies. A distinct elliptical precipitin arc results for each protein detectable by the antisera.
D007525 Isoelectric Focusing Electrophoresis in which a pH gradient is established in a gel medium and proteins migrate until they reach the site (or focus) at which the pH is equal to their isoelectric point. Electrofocusing,Focusing, Isoelectric
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000918 Antibody Specificity The property of antibodies which enables them to react with some ANTIGENIC DETERMINANTS and not with others. Specificity is dependent on chemical composition, physical forces, and molecular structure at the binding site. Antibody Specificities,Specificities, Antibody,Specificity, Antibody
D001323 Autoantibodies Antibodies that react with self-antigens (AUTOANTIGENS) of the organism that produced them. Autoantibody
D013954 Thyroglobulin

Related Publications

L Nye, and I M Roitt
December 1981, Journal of biochemical and biophysical methods,
L Nye, and I M Roitt
July 1987, Journal of virological methods,
L Nye, and I M Roitt
January 1989, Journal of biological regulators and homeostatic agents,
L Nye, and I M Roitt
January 1994, Bulletin de la Societe des sciences medicales du Grand-Duche de Luxembourg,
Copied contents to your clipboard!