stripping and western blot?
khatipovNO at NOuchicago.edu
Fri Aug 30 11:31:53 EST 2002
"Schwabe, TME" <tatjana.schwabe at ruhr-uni-bochum.de> wrote in message
news:3D6F9974.9010707 at ruhr-uni-bochum.de...
> > Seems interesting. I never heard of using low pH in Westerns, although
> > widely used elsewhere. The only thing I wonder is how you make glycine
> > buffer pH 1.9, when none of the conventional pH meters can accurately
> > measure such a value? Could you post you method or the source here?
> > Emir
> Oops, I think you caught me on something there...
> Basically we just titrate with HCl to what our pH-meter calls pH 1.9. I
> just checked with one of the little pH-indicator strips and the pH is
> somewhere between 1 and 2. I'd say it doesn't matter so much. I know
> that in other protocols you elute AB with pH 3, so this should probably
> suffice in Westerns as well.
> I don't have a source for this protocol though, sorry.
One can always buy ready-made glycine 1.9 or whatever from chemical
suppliers. I use glycine 1.5-2.5 in BIAcore experiments and buy the buffers
from BIAcore, though it is quite expensive (but !guaranteed quality! :-))
... Still wonder how THEY measure pH1.9. Guess they might be using
precalculated volumes and quantities instead.
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