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protein purification


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#1 anonymous

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Posted 29 January 2001 - 10:00 PM

You can use an enterokinase immobilized on Sepharose. Search the Sigma catalog for "Enterokinase". I just did it online, Sigma calls the product "CleanCleave". They alsohave a product called "Enterokinase Removal Kit" product code PRKE. Seems to be an immobilizedantibody against enterokinase. Either of these would likely do the job you are asking about.

#2 anonymous

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Posted 29 January 2001 - 10:00 PM

hi!i have used the vector pRSETB for cloning one of the genes.my protein will ultimately have a histidine tag and an enterokinase clevage site.my question is,after isolating the protein of interest and using enterokinase to seperate the his tag and the protein of interest, how do i get rid of the enterokinase and then get the protein of interest.am not interested in going for the kit which will get rid of enterokinase, provided by invitrogen.any answer?

#3 anonymous

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Posted 02 February 2001 - 10:00 PM

You could also use dialysis. After cleaving the linker off, select dialysis tubing which will let your smaller molecular weight linker dialyse out, while keeping your protein inside. I've done this with a protein tagged with a His tag also but with a thrombin cleavage site. Run your results on a SDS-PAGE against the uncleaved protein.

#4 anonymous

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Posted 02 February 2001 - 10:00 PM

You could also use dialysis. After cleaving the linker off, select dialysis tubing which will let your smaller molecular weight linker dialyse out, while keeping your protein inside. I've done this with a protein tagged with a His tag also but with a thrombin cleavage site. Run your results on a SDS-PAGE against the uncleaved protein.




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