I tried to clean my Superdex 200 AKTA gel filtration column after an event of protein sample aggregation on the column during gel filtration. Unfortunately, the column seems to be literally blocked, even at the lowest possible 0.05 ml/min flow rate I cannot go further without reaching the pressure limit of 1.5 MPa. This final failure happened after about 8-9 ml of 0.5 M NaOH solution entered the column. I suppose it is not good to leave the column in these conditions (pH 10) overnight, therefore I would like to connect the column in reverse flow but I cannot find a procedure how to do this. Thank you for your kind help and supportive ideas- even a link to useful contents would be of great help.
If so, you don't have to reconnect the column!
Simply go to manual control-->Flowpath-->Flow direction and select upflow
Hope nobody messed up the tubing connectors on your system.
If this doesn't work, open tubing connector at the bottom of the column to check if the column really is the problem.
Best,
Kai
Hello gergely,
You should open your column and change the filter at the top. It should sort part of your pressure problem. Usually that is were blockage happen. When finish do a proper cleaning procedure.
Good luck
bertrand
Hi Gegerly! I was wondering if you could eventually fix the issue with your gel filtration column. If yes, then how? Or did you use the column in reverse orientation for purification?
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