[Protein-analysis] Protein purification under denaturing contitions

Julio Cortazar juliocortazar at o2.pl
Thu Jul 27 23:23:24 EST 2006


Hi there

Is is true that protein purification under denaturing contitions may be
done in room temperature? I'm puryfing my protein from inclusion bodies
using urea solutions and nickel-agarose column. In 4Cdeg urea in my
buffers starts to crystalize! If I perform lysis of the cell in room
temp thus I think all the proteins (also proteases?) are denatured, am
I right? Next step wolud be refolding of the protein in multistep
dialysis, so I will have to make it partially in 20 partially in 4C.

Julio



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