Transfer of proteins and transfer buffer formulations...
Bernard Murray, PhD
spam at 127.0.0.1
Mon Feb 7 19:40:06 EST 2000
<Pine.A22.214.171.12402052249050.91100-100000 at acs6.acs.ucalgary.ca>, Neal
Robert Melvin <nrmelvin at ucalgary.ca> wrote:
> Can anyone recommend a transfer buffer composition that will more
> efficiently transfer proteins between 120 and 180kDa?? I'm currently using
> a transfer buffer that has 0.1% SDS and 20% methanol. I've tried using
> more SDS to facilitate transfer, but when I stain my gel with Coomassie, I
> still get the same higher molecular weight bands in the gel after
> transfer... would altering the methanol concentration help??
Its a trade-off as putting SDS in the buffer increases the efficiency
of extraction of proteins from the gel but decreases the efficiency
of sticking to the membrane. Methanol has the reverse effect.
Dropping the methanol concentration would probably help.
This is also one of the situation where I would recommend using
a semi-dry transfer unit.
Bernard P. Murray, PhD
bpmurray at cgl . ucsf . edu
Department of Cellular & Molecular Pharmacology, UCSF
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