EBlot

rowland@panda.uchc.edu
Mon, 23 Nov 1998 15:27:25 -0500

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Hi,
I would like to know if anyone has experience with electroblotting, and
western detection of very small proteins (2-8 kDa). I am running 16.5%T,
6%C tris-tricine gels with a 10%T spacer gel. These give reasonable
resolution. I am having trouble getting reproducible transfer of these
proteins (standard tris/glycine/20% methanol transfer buffer) and am
getting no signal on western blots. A control protein of 10 kD gives a
positive on the western with the same antibody (and the small proteins
contain the epitope to which that antibody was raised). I have tried small
pore (0.1 um) nitrocellulose and PVDF membranes. I would appreciate any
suggestions.

Thank you, Susan

Dr Susan Rowland
Department of Microbiology
University of Connecticut Health Center
Farmington, CT, 06030, USA

Email: rowland@panda.uchc.edu
FAX: 1-860-679-1239
PH: 1-860-679-2203

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