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co-eluted protein with his-tag protein


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#1 alinud

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Posted 23 July 2012 - 11:51 PM

Hello,
I have recently expressed a protein in BL21, about 31 kDa with Histag (Flexi system, Promega).  I tried to purify the protein using  MagneHis Protein Purification System (Promega). I find that my elute contains most protein in the host cell. I’ve tried increasing the NaCl conc in the wash buffer, but the problem still exists.
Any suggestion what’s happening? Thanks for help.

#2 Inmost sun

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Posted 24 July 2012 - 08:13 AM

did you check if your protein of interest was really overexpressed? If so, was it prominent in the elutions?




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