I would like to do a native gel on a small (9 kDa) basic (pI-9.5) protein, and we will be looking to resolve different isoforms that have different PTM's. These PTM's will alter the pI significantly (by 1-2 pH units), so we would need an electrophoresis system that covers a range of pH's, like 6-9. I read that acetic-acid urea gels are good for basic proteins, but could that be used in this case? Any ideas would be appreciated!
I support the idea of Vivek Hamse, in which you should use acetic acid gel at the pH between 5-6. This will allow your protein to migrate through the gel without any effect of the acid on the activity of your enzyme "especially if you would like to detect the enzyme on the gel.
I never tried it, but have you thought about "Blue Native PAGE"? Binding of coomassie gives your protein a negative charge.
Thanks for your help everyone! I will give this a try. Vivek, my email is [email protected]. I'd like to see which protocol you use.
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