Problem with Acid gels for basic proteins

Sonya Clark szsclark at
Sat Aug 19 19:36:47 EST 1995

Dear Netters,

I am wondering if you could advise me on what I can do to improve my acid
gels for basic proteins.  I am following the recipes of Reisfeld et al. 
which uses a KOH/Acetic Acid buffer, Stacking gel at pH 6.8, Running gel
at pH 4.3 and beta-alanine/Acetic acid running buffer at pH 4.5.  I am 
including 8M urea in the running gel and am correcting my buffer pH so 
that the final pH (with urea) is 4.3.  My protein of interest has a pI of 
10.1 and runs into the gels O.K., but looks like a comet rather than a 
band.  The protein is solubilized inclusion bodies in 8M urea, and looks 
great on standard SDS-PAGE gels, but I need to be able to run it on a 
native gel for various experiments I'm planning.  Any suggestions on how 
to improve my gels?  Any thoughts are appreciated! Thanks. 

Dr Sonya A. Clark
saclark at
Section of Plant Biology
University of California
Davis CA 95616

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