I have a home made procainamide sepharose 4B column that completely filled by contaminant proteins. Can I use of EDTA as cleaning agent? Is it possible the EDTA destroy the procainamide?
I don't think EDTA will damage the column, but I also would be surprised if it helped clean it. Try high salt (1M NaCl) and low pH (pH 2) washes, also.
I am not sure how it works with EDTA,..
but we use Sephadex columns in our lab.
And cleaning process we use is:
overnight shaking incubation of column(with clean buffer) at higher temperature than optimal temperature of protein [say 50-60 degrees for proteins stable @ room temperature],.. after that the buffer is drained and fresh clean buffer is run through column
You can use 2% EDTA for cleanliness, or use hot water treatment of packing material, the best way to clean the matrix is take it out suspend in PBS buffer, centrifuge at 5000 rpm three times and repack it.
thank all of you
Adam: for Amersham nickle column stripping, they have a protocol that used EDTA as cleaning agent.
EDTA is used to strip nickel or other metals off of IMAC columns because it binds more tightly to the metal than the column does. Any His-tagged proteins remaining on the column due to their interactions with the bound metal will be stripped off at the same time. If metal cations are important for the interaction of proteins with procainamide, then EDTA may help clean them off your column. Otherwise, not.
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