Native gel problems

JMORELLI at MAINE.MAINE.EDU JMORELLI at MAINE.MAINE.EDU
Wed Dec 7 13:08:27 EST 1994


I am trying to run a native (no SDS) acrylamide gel on some basic
proteins. (myoglobin, hemoglobin)  I hav so far not had much luck.
I notice that my prestained size standards run very different from
their usual size separation, one standard will not even run into the
gel.  I know that the pI of the proteins is a factor, but
I can't seem to find information on how to adjust the sample buffers
and gel running buffers to compensate for protein charge.  I assume
that I need to make the buffers more alkaline than the protein
pI to impart a net negative charge.  I have not found any
references where the pH of the buffers is changed in any way from
the normal SDS-PAGE gel.  Only SDS and DTT are omitted.
My question is:  Does anyone know of a protocol, or a reference
for native gel separation of basic proteins?  I would appreciate any
help you can give me.  Our Email computer may not be working properly,
so please post directly to this newsgroup.
Thanks,
Jim



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