Protein A column still okay if it was let dried? - (Feb/01/2007 )
I was purifying some antibody using a protein A column. (Biorad cat#732-2022) It is made of protein A linked to agarose beads.
At the last step, when I am regenerating the column, the regeneration column was completely dried.
I immidiately added more regeneration buffer and allowed it to enter the column.
My question is: does anyone know if this column is still good to use?
your column will probably be okay. it does not appear that you allowed it to stay dry for an extended period of time.
that being said... it would probably be best if you unpacked, defined the gel and repacked the column. but it should still work okay if you don't.
that being said... it would probably be best if you unpacked, defined the gel and repacked the column. but it should still work okay if you don't.
What is the procedure for repacking?
i just looked at the product description at the bio-rad website. it is probably not worth the effort to repack the column. just use it as it is (get rid of any air bubbles for best efficiency).
i just looked at the product description at the bio-rad website. it is probably not worth the effort to repack the column. just use it as it is (get rid of any air bubbles for best efficiency).
I was told by a colleague to run the column at a very high flow rate in order to "re-pack" the column.
Do you think this is a good idea? Will it hurt the column?
HI!
I think it is not a good idea. YoU can damage agarose particles I mean it will appear a small particles and as a result increasing back pressure will occur. Repack the column. It is not very difficult techniq (of course if it is not prepacked Amersham HI-TRAP) 
you can flow buffer in from the bottom. this will drive air out of the gel. it does not need to be run fast, in fact, it is better to run slowly.