A modified Western blot protocol for enhanced sensitivity in the detection of a membrane protein.

Kaur, Jaspreet and Bachhawat, Anand K (2009) A modified Western blot protocol for enhanced sensitivity in the detection of a membrane protein. Analytical biochemistry, 384 (2). pp. 348-9. ISSN 1096-0309

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Abstract

Membrane proteins, owing to their highly hydrophobic nature, have always posed a daunting challenge to biochemists and structural biologists working on the characterization of these "naughty" proteins. Here we describe a problem that we encountered in the immunodetection of a hemagglutinin (HA) epitope-tagged membrane protein, Hgt1p (high-affinity glutathione transporter from the yeast Saccharomyces cerevisiae), for which little or no signal was observed on the blots with monoclonal antibody, on following the standard Western blot protocol. The introduction of a single step that involved posttransfer incubation of the blots with sodium dodecyl sulfate (SDS)/beta-mercaptoethanol solution at 55 degrees C for 15 min enabled us to detect a strong, stable, and reproducible signal for the membrane protein.

Item Type:	Article
Additional Information:	Copyright of this article belongs to Elsevier Science
Subjects:	Q Science > QD Chemistry
Depositing User:	Dr. K.P.S.Sengar
Date Deposited:	08 Dec 2011 19:37
Last Modified:	08 Dec 2011 19:37
URI:	http://crdd.osdd.net/open/id/eprint/577

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