migration of a non denaturant PAGE
no-sp at m.org
Wed Jul 11 02:30:08 EST 2001
Try a normal SDS PAGE, just leave out the SDS. Compare the pH in the buffer
to the pI's and figure out whether your protein will run into to gel or out
of the well. In the latter case, reverse the current. In this case you
should also use another tracking dye, e.g. methyl green.
You could also add some urea, making the gel denaturing without masking the
charge differences of the two proteins. Perhaps you could use acetic
acid-urea PAGE (see medline).
What about isoelectric focusing?
"Stefi~" <xxxgifalone at tin.it> wrote in message
news:gYH27.1615$2v6.44761 at news.infostrada.it...
> I have two proteins with very similar mol weight (and very similar (but
> identical!) pI. When I do a SDS-PAGE and blotting I find bands at the same
> position. As my antibodies are not very specific, I'd like to use blotting
> on a non denaturant PAGE, hoping I'll have different migration. What do u
> gifalone at tin.it
> ICQ 110419911
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