oxidation of SS bridges

Vennie ossipow at sc2a.unige.ch
Wed Mar 29 03:48:47 EST 1995


In article <3l8hse$sk at lyra.csx.cam.ac.uk>, cdw1000 at mole.bio.cam.ac.uk (Christopher Walentas (Bioc)) says:
>
>Hi.
>
>I was wondering if anyone could share their favorite protocol for
>the oxidising of disulhpide bridges.  I've engineered in a couple
>to stabilise my enzyme, and they're there as far as a DTNB assay 
>under denaturing conditions is concerned, but the buggers don't
>seem to want to form.  Perhaps they never will, but I'd like to
>exhaust other options first.  Folki have mentioned CuSO4, K3Fe(CN)6
>and iodosobenzoic acid, but I've got to track down some protocols
>to get an idea of final concentrations, pH, etc...  ANY help on
>the subject would be appreciated!
>
>Cheers in advance!
>
>Tripp
>
>-------------------------------------------------------------------------------
>C. D. Walentas
>Cambridge Centre for Molecular Recognition,
>University of Cambridge,
>Cambridge, England CB2 1QW
>
>Phone:  0223 333 662
>Fax:    0223 333 345
>email:  c.d.walentas at bioc.cam.ac.uk
>
I also wanted to oxidize two monomers to form a covalently linked dimer.
I tried different solution as H2O2, which gave good and clean results 
(send me a email if you want the reference), Zn or Cu solutions, but
the cleanest and quite efficient in my case was incubating my proteins
 overnight in a oxygene atmosphere with shaking.

 Good luck Vennie




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