Hi All,
I'm regenerating TALON beads in bulk for the lab. As I've done with Nickel beads for years, I washed with water, eluted all the protein, removed imidazole, and then added 0.2M EDTA pH 8.0 (as per the instruction manual). I saw some of the pink color get removed as the EDTA worked through the column, but there was still a lot of pink left-over. (For reference, all of the Nickel was stripped off of the nickel beads I was regenerating in a separate column at the same time, so the EDTA seems fine and the Ni beads turned white).
I tried again with 0.5M EDTA pH 8.0 and saw no change. So, thinking maybe some protein or imidazole was still attached to the cobalt, I washed with 250mM imidazole, water, and then the recommended 20mM MES pH 5.0, 0.1M NaCl buffer to remove imidazole. Then water again, then 0.5M EDTA pH 8.0 and saw no change.
Is the pH of the EDTA too high? I saw somewhere that someone said the gel itself is pink so I shouldn't worry. Can anyone confirm this?
I've been trying all of the washes in the manual to no avail.
My plan for now is to just recharge anyways as I know some of the cobalt was stripped.
Thank you!
Hi Jillian Breault
that is a very interesting observation. I have just checked the protocol of the Talon resin of Takarabio. On page 10 there is a small description how they recommend regeneration. They just wash it with MES buffer. So it seems that it is just not possible to strip the Cobalt ions from the resin. Your observation fits that. The resin is to complexed and the Cobalt ions are retained. We have a different resin which is Co-NTA based. It is a very similar technique and we use this regeneration protocol (https://cube-biotech.com/washing-and-regenerating-his-affinity-magbeads-and-agarose-resin).
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