? Difficulty with Factor Xa cleavage ?

[Jongrak Kittiworakarn]

Dear All:

I expressed my peptide of interest (one is of 6 kDa) with GST fusion
vector,pGEX-3x.  The protein was expressed in the insoluble form, and
refolded into soluble protein.
I can detect very low GST activity in the refolded protein, but the
protein did not bind to s-hexylglutathione agarose. Then I tried to
cleave my peptide out from the GST part,but I could not. I also tried
the cleavage in 0.5M and 0.25M urea, but it did not work as well.
It would be prefered not to use any detergent in my excperiment.

Thank you very much in advance for any suggestion.

Jongrak



-----------------------------------Mr. Jongrak Kittiworakarn

g3937506 at student.mahidol.ac.th

     Inst. of Science and Technology for Research and Development
     Mahidol University (Salaya Campus)
     Salaya, Nokornprathom, THAILAND.
     Tel. 001-66-2-441-9003-7 ext.1277,1278,1279
     Fax. 001-66-2-441-9906, 001-66-2-441-1013
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