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Protein purification. How to make function of recombinant protein?


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#1 mylovesci

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Posted 11 February 2011 - 11:11 AM

I am senior student in bachelor degree. I have a problem. I purifind recombinant protein with Ni-chromatography in order to treat cell. However, i don' t know that the cell was died by my reconbinant protein or imidazole because imidazole may be affect the cell. Will I do it, if I does change into water or any compound which does not damage a cell? Could someone suggestme.
p.s. If you have paper, May you tell me too. Thank you for your kind.

#2 mdfenko

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Posted 11 February 2011 - 11:46 AM

did you also use urea when purifying the protein from the column?

if not then you can eliminate the imidazole by dialyzing the protein against a more compatible buffer.

if you used urea then you will have to refold the protein by stepwise dialysis to remove urea (and you can remove imidazole at the same time).
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#3 mylovesci

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Posted 12 February 2011 - 07:42 AM

To mdfenko
Thank for your suggesion. I use urea ,so i will try stepwise dialize on monday

Edited by mylovesci, 12 February 2011 - 07:46 AM.





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