HELP: PROTEIN SOLUBILIZATION

[T. J. Murphy]

Hello,
I am studying an integral plasma membrane protein (very hydrophobic) that we suspect
is forming an insoluble multimeric complex when prepared for SDS-PAGE
(sample loading buffer=4%SDS, 2-ME, pH 6.8).  The results are the same whether or
not the samples are boiled.  Immunoreactivity on Westerns appears
to be retained in the wells of the stacking gel.  We would like to
get this protein into the separating gel for analysis.

Does anybody have any specific protocols for treating a plasma membrane
fraction sample to overcome this problem?  I would be grateful for any
advice, preferably by e-mail direct.  I can later summarize the suggestions
for the group. 

medtjm at bimcore.emory.edu
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TJ Murphy
Asst. Professor	      
Dept of Pharmacology
Emory University School of Medicine