Problem with Acid gels for basic proteins

Sonya Clark szsclark at chip.ucdavis.edu
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 ucdavis.edu
Section of Plant Biology
University of California
Davis CA 95616
----------------------



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