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,


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