Transfer of proteins and transfer buffer formulations...

Dima Klenchin klenchin at facstaff.REMOVE_TO_REPLY.wisc.edu
Sun Feb 6 13:40:19 EST 2000


: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??
:

Yes, decrease methanol to 10%. 

Alternatively, use CAPS buffer based transfer buffer _without_ SDS
(high pH --> higher charge --> better transfer).

        - Dima




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