Re: [ccp4bb] How could I Extract Iron from Protein?

[Roger Rowlett]

I would try dialyzing against a solution with 1,10-phenanthroline, 1,10-phenanthroline-2-carboxylate, or pyridine-2,6-dicarboxylate. Removal of metals from proteins is often not just dissociative, but requires the associative interaction of a chelating agent. Which one works is often empirical. Cheers. On 5/11/2010 11:11 AM, Wenguang LIANG wrote: Dear all,   I have a protein which binds iron with two D and two H as active site. I have tried to extract the iron by dialysis. First, 20mM tris, pH7.5, 150mM NaCl, 20EDTA, 10mM Na2S2O3, O/N. Then, followed by dialysis with 20mM tris, pH7.5, 150mM NaCl, 1EDTA O/N to remove the EDTA and Na2S2O3. However, after dialysis, the density of the Iron is still in the protein.   Could anybody please give me some suggestion on how to extract Iron out of the protein? Or I can just use Chelex-100 to extract it instead of Iron.   Thank you very much for help,   Best wishes,   Vinson Liang   -- Roger S. Rowlett Professor Department of Chemistry Colgate University 13 Oak Drive Hamilton, NY 13346 tel: (315)-228-7245 ofc: (315)-228-7395 fax: (315)-228-7935 email: rrowl...@colgate.edu