Running Polyacrylamide gel in acidic conditions - Adicid denaturing PAGE (with or without SD?) (Jun/18/2009 )
I am trying to run a highly basic proein (pI >9.5) on a polyacrylamide gel. Apparently I load a huge amounts of protein (around 10-20 micrograms in one well) on gel, i see vague bands. I have used the acidic conditions for gel in native state (non-denaturing gel) and I see good bands on gel. I am not sure, if that works in presence of SDS + acidic conditions, Any body has ever tried that and is there any protocol, that i can use for running my protein in denaturing conditions in acidic polyacrylamide gel.
Thanks in advance
if you want to run under denaturing conditions then just run a standard sds gel. the sds will impart a net negative charge to the protein at the pH of the gel and running buffers.
you could run the acid gel with urea.