J Biochem Biophys Methods. 1990 Jun;21(1):65-8.

Elimination of non-specific binding in western blots from non-reducing gels.

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Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.

Abstract

The reaction of some antibodies with Western blots of protein shows strong non-specific binding especially at a region that corresponds to about 70-90 kDa. This binding is independent of protein concentration. Further analysis indicated that the factor responsible for the non-specific binding is 2-mercaptoethanol in the gel sample buffer. Gel electrophoresis of total tissue homogenates in the absence of this reducing agent resulted in dramatic elimination of the non-specific background binding without affecting the mobility of the two proteins we studied.

PMID:: 2212467; [PubMed - indexed for MEDLINE]

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