FPLC Column Cleaning Protocol?

noone noone at cancer.bham.ac.uk
Thu Jul 30 07:20:52 EST 1998


> 
> Seems to be more a mechanical problem. Try to check tubings and filters. A
> protein/DNA etc.- plugged column should easily be cleaned by NaOH up to 2M,
> HCl 1M and Urea (don't forget to run a normal buffer in between).
> 


And do not only check the filter of your column but also the filter behind
the mixer (check if the high backpressure persists if you run the machine
without a column). In extreme situations , you may treat your column with
pepsin in a low pH-buffer (I know, proteases and columns are a scary
thought, but pepsin should be pretty inactive at neutral pH) to get rid of
clogged-up protein.
Run your column upside down to loosen the compressed material a bit while
you clean it

Hope this helps,

Peter



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