Blocked column

Alison Richmond alison.richmond at strath.ac.uk
Thu Apr 4 12:15:15 EST 2002


Hi,

I have just loaded on all my protein on to a DEAE-Sephacel column and the
column now appears to be blocked, as some of the tubing has been forced
apart due to back-pressure.  My protein is coloured and I can see it!! But
not sure exactly how to recover it.  Should I be able to resuspend the
column material, mix with a high salt buffer (which I  would normally use to
elute the protein) and then by centrifugation recover the protein in the
supernatant, repeating the proceedure until the column matrix appears white
again?

Any help and advice much appreciated,

Alison


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