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Immunoprecipitation-Western Blot for Proteins of Low Abundance

  • Edward P. Trieu
  • Joanne K. Gross
  • Ira N. TargoffEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 536)

Summary

Combining the procedures of immunoprecipitation and immunoblotting can help to overcome some of the limitations of each separate procedure. Immunoblotting can identify immunoprecipitated proteins more specifically and with higher sensitivity than nonspecific protein stains or autoradiography. Immunoprecipitation can enrich proteins of interest to improve sensitivity for detection when compared with immunoblotting of whole cell extracts. Recently, immunoprecipitation-blotting helped to characterize a new autoantibody, anti-p155, and to test for the presence of the autoantibody in patient sera to study its clinical associations. The procedure for immunoprecipitation-blotting, with specific reference to this autoantibody test (“reverse” immunoprecpitation-blotting), is reported here in detail.

Key words

Immunoprecipitation Immunoblot (western blot) Autoantibody Autoantigen Dimethyl pimelimidate 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Edward P. Trieu
    • 1
  • Joanne K. Gross
    • 1
  • Ira N. Targoff
    • 1
    Email author
  1. 1.Arthritis and immunology ProgramOklahoma Medical Research FoundationOklahoma CityUSA

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