Characterization of metabolically labelled antigen following immunoprecipitation with antibody bound on ELISA plates.
Immunoprecipitation is a powerful technique for the immunochemical characterization of antigens. In combination with sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) a number of features, e.g., presence of antigen, rate of synthesis, relative molecular weight of the polypeptide chain or post-translational modifications can be determined. Four different steps are basically involved in the immunoprecipitation procedure: (1) metabolic labelling of the antigen by incubation of viable cells with a radioactive precursor, (2) harvesting of the labelled antigen from the cells by lysis or in the case of secretory proteins from the supernatant, (3) formation and (4) purification of antibody-antigen complexes. The last step relies on secondary agents which bind to the antibody.
von Brunn
A
A
Max-von-Pettenkofer-Institut, Munich, Germany.
Reichhuber
C
C
D000818:Animals; D000936:Antigen-Antibody Complex / Q000302:isolation & purification; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005347:Fibroblasts; D011817:Rabbits; D015531:Radioimmunoprecipitation Assay / Q000379:methods*; D011864:Radioimmunosorbent Test
D000936:Antigen-Antibody Complex