Western blot and isoelectric point - (Jan/17/2012 )
Continuing with my previous problem. I found that all my proteins have pI
Methanol strips the SDS from proteins and proteins are finally transferred to membrane - I was wondering whether really the pI will affect the SDS-PAGE followed by western blot OR the pI is imp only if use the proteins in native form ?
Any one faced problems with proteins with high pI
with sds present pI is not important. pI is important for native separations.
you can try a CAPS buffer at pH 11 for the transfer.
i am attaching a book from millipore and you can download a manual from ge healthcare from this link:
western blotting - priciples and methods