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inactive enzyme after purification with Ni-TED
Recombinant Protein Forum
Recombinant Protein Forum. Discuss purification, expression, and isolation of recombinant and cloned proteins here.
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05-02-2008, 08:14 AM
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 inactive enzyme after purification with Ni-TED Hi everybody I expressed a 54 kDa protein in E.coli. Before the purification with Ni-TED the enzyme is active. After the purification there is no activity left. What can be the reason? It is a metalloenzyme and needs Mn 2+ for its activity. Could it be that the imidazole in the buffers while purification is the reason? How can I then reactivate the enzyme? Or do the Mn Ions bind to the column while purification? Or what else could be the problem?         |
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