clean-up of recombinant protein before assay

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wildaisy
wildaisy's picture
clean-up of recombinant protein before assay

I expressed and purified recombinant proteins and added them into cell culture to test their effects on cell growth, but contaminated cell culture. Seems the origin of the contamination is from the recombinant protein. Should I clean up my protein before using them? Any way to clean it up?Thanks.

Ivan Delgado
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What I would do is filter sterilize your protein solution through a 0.22 filter. This should remove any bacteria present while at the same time be as gentle as possible on your protein's activity. If this does not work you could go the extra mile and incubate your protein solution with your favorite anti-bacterial/-fungal and then remove it by filtration.

RLS
RLS's picture
 You can also try using a gel

 You can also try using a gel filtration column (something like a superdex 75 or 200), which will not only clean up your protein of other protein contaminants, but you won't have any bacteria in there either. You can use very gentle conditions on these columns, but it's a little longer of a process than simply filtering your protein of interest, as Ivan suggested.

wildaisy
wildaisy's picture
thanks, Ivan and RLS. I am

thanks, Ivan and RLS. I am actually trying the 0.22um filter, hope that will work :)