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.
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Protein purification. How to make function of recombinant protein?
Started by mylovesci, Feb 11 2011 11:11 AM
2 replies to this topic
#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).
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
Thank for your suggesion. I use urea ,so i will try stepwise dialize on monday
Edited by mylovesci, 12 February 2011 - 07:46 AM.
