I had exactly the same problem. You should switch to Talon matrix
(Clontech). The cobalt ion is bound more tightly so stripping is
dramatically reduced. Furthermore, the Talon matrix is more specific
(then Ni Sepharose) for your His-tagged protein, so you have less
contaminants. I load two to three liters of culture medium directly on a
20 mL Talon column. This works very good in my hands.
Good luck,
Kenneth
Oganesyan, Vaheh schreef:
Dear All,
When mammalian cell culture is being loaded to GE HisTrap resin Ni ions are being stripped off the resin, at least in my hands. Did any of you have similar experience and if so what kind of work-around was found?
Volume is fairly large (3L) and concentration/dialysis have proven to cause
loss of desired protein.
Please share your positive experience.
Thank you for your time.
Vaheh
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