Finding a suitable denaturing agent |
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Hi, everyone
I got some problems in denaturing the endogenous collagenase of planarian extract. I had try to use 8 M of urea and complete protease inhibitor for extraction, but I still cannot inhibit the collagenase activity.
what another denaturing agent that I can used to solve this problem? Does guanidine - HCl is a good choice?
thank you v.much~
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Replies
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| cindyfunnybaby said: | | Hi, everyone I got some problems in denaturing the endogenous collagenase of planarian extract. I had try to use 8 M of urea and complete protease inhibitor for extraction, but I still cannot inhibit the collagenase activity. what another denaturing agent that I can used to solve this problem? Does guanidine - HCl is a good choice? thank you v.much~ |
GuHCl at 6M is also worth trying. You can also try heating your sample in the 8M Urea or 6 M GuHCl. Does collagenase have disulfide bonds? Are you adding reductant, DTT or BME? I have not used GuHCl at higher than 6M. Anyone else?
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Im pretty sure GuHCL will denature anything at 4M. Will try to find a reference.
"Everything should be made as simple as possible, but no simpler."
-- Einstein
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