Does anyone have experience with Elution of Fe-S protein from Ni-NTA column. My problem is that protein is well bind in Ni-NTA column but when I elute with imiadazole the reddish brown colored region containing Fe-S protein is not eluted even though I tried 100-1000mM imidazole.
Marc Birringer
Dear Atul,
very tricky! I assume the sulfuratoms coordinate with the Ni-Atoms. May add some mercaptoethanol in addition to imidazol. That could separate the S-Ni-Bond but also change the redox state of your Fe-S-cluster.
Best
Marc
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