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Can J Microbiol. 2007 Apr;53(4):526-32.

Conditions that allow for effective transfer of membrane proteins onto nitrocellulose membrane in Western blots.

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  • 1Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON, Canada.

Abstract

A major hurdle in characterizing bacterial membrane proteins by Western blotting is the ineffectiveness of transferring these proteins from sodium dodecyl sulfate -- polyacrylamide gel electrophoresis (SDS-PAGE) gel onto nitrocellulose membrane, using standard Western blot buffers and electrophoretic conditions. In this study, we compared a number of modified Western blotting buffers and arrived at a composition designated as the SDS-PAGE-Urea Lysis buffer. The use of this buffer and specific conditions allowed the reproducible transfer of highly hydrophobic bacterial membrane proteins with 2-12 transmembrane-spanning segments as well as soluble proteins onto nitrocellulose membranes. This method should be broadly applicable for immunochemical studies of other membrane proteins.

PMID:
17612609
[PubMed - indexed for MEDLINE]
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