Concentration of dilute proteins for SDS-PAGE
fortinfl at ERE.UMontreal.CA
Sat May 4 12:14:28 EST 1996
Joe Stains <JPS144 at psuvm.psu.edu> writes:
>Does anyone know of any efficient way of concentrating large volumes of
>a dilute protein (several mls) for use with SDS-PAGE? I have a couple
>of ideas on some techniques, but would be interested in anything anyone
>else may have, as they all have a lot of pros and cons!
>Thanks in advance
What I usually do to concentrate my proteins, is to precipitate them with 3 volumes of ice-cold 100% acetone. Leave on ice for 10 minutes, then spin 10 minutes at 10 000 g. Discard the supernatant. Brief wash with 80 % acetone. Spin again and let dry the pellet for 15-20 minutes. Resuspend your proteins in the volume of your choice with PBS or else. You can also use those concentrators from the company amicon or everything like that, but it can take several hours to obtain what you whant in volume. Ho
pe it will help you.
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