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Nonelectrophoretic Bidirectional Transfer of a Single SDS-PAGE Gel with Multiple Antigens to Obtain 12 Immunoblots

  • Biji T. KurienEmail author
  • R. Hal Scofield
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 536)

Summary

Protein blotting is an invaluable technique in immunology to detect and characterize proteins of low abundance. Proteins resolved on sodium dodecyl sulfate (SDS) polyacrylamide gels are normally transferred electrophoretically to adsorbent membranes such as nitrocellulose or polyvinylidene diflouride membranes. Here, we describe the nonelectrophroretic transfer of the Ro 60 (or SSA) autoantigen, 220- and 240-kD spectrin antigens, and prestained molecular weight standards from SDS polyacrylamide gels to obtain up to 12 immunoblots from a single gel and multiple sera.

Key words

Nonelectrophoretic transfer Immunoblots Bidirectional transfer Nitrocellulose ­membrane Autoantibodies Autoantigens 

Notes

Acknowledgement

This work was supported by NIH grant ARO1844 and Oklahoma Center for the Advancement of Science and Technology to RHS.

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Copyright information

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

Authors and Affiliations

  1. 1.Oklahoma Medical Research FoundationOklahoma CityUSA

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