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GE Q High Performance Resin question
Trying out the GE High Performance Q Resin and wondering how others experiences with this resin go.
Performing a protein purification under denaturing conditions in 6M Urea. First step is IMAC chromatography second step is Q.
So protein is going on to the column in 20 mM Tris, 6M Urea pH 8.0.
Thing is I am only getting 50% yield. I'd be fine with this if I could find the rest of the protein. Evaluated each peak (only 3), flow through, washes, and boiled the resin and that all adds up to ~10%, protein of interest is 50%, so where the hell did the other 40% go?
So if anyone has some troubleshooting techniques or conditioning for Q that would be great. I'm starting there but of course it could be anything including my BCA assay.
|high , performance , question , resin|
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