Hi
I am purify a his tagged protein expressed in E coli Bl 21DE3. But after gel filtration there are still contamination of different size of protein. I tried Ion exchange chromatography but it did not worked. Is there any thing i can do so that the unexpected protein will not be expressed or how to purify that one
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2 replies to this topic
#2
BioMiha
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Posted 15 February 2011 - 02:01 AM
Usually His-tagged proteins are purified using a Ni-affinity column. You can play around with different concentrations of imidazole to try to achieve best purity.
#3
scolix
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Posted 15 February 2011 - 11:25 AM
try a different system like from HEK cells. sometimes, its difficult to get a pure protein prep from E.Coli.
but is the contamination a real problem for your exp.?
but is the contamination a real problem for your exp.?
