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how to deal with protein precipitates during dialysis

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#1 dachshund



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Posted 04 November 2009 - 05:18 PM

Hi all:

I am now trying to purify my His-tagged protein. Some of the protein precipitated during dialysis, my next step is to cleave the protein using EKmax protease. It seemed like that the protease did not digest the precipitated proteins. What should I do with those precipitates? Any suggestion is appreciated!

#2 pesji



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Posted 05 November 2009 - 05:08 AM

Can't you do the digest before the dialysis ? Usually the precipite appears when going from Urea to native buffer (such as PBS) you might be able to solubilize again the precipitated protein in Urea (try 1M)

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