Hi,
you may also check things like chemical degradation in SDS buffer as part of
the analysis. Esspecially your degradation pattern is very much constant
throughout your whole purification procedure.
Christian
Am Mittwoch 15 Februar 2012 14:09:19 schrieb Sivasankar Putta:
> Dear All,
>
> Can anybody suggest the tricks and trades of stabilizing a 133 kDa (multi
> domain) DNA binding protein, that we are expressing at 18 degree Centigrade
> in* E. Coli.* The protein appears to degrade during purification; we have
> protease inhibitor cocktail (in the lysis buffer) as well as 2 mM PMSF, 1
> mM EDTA and 1mM DTT throughout during purification ( right from lysis
> stage). We handle the protein at 4 degree Centigrade.
>
> Can you please suggest what precautions we can try to avoid such
> degradation ?
>
> Please find the attached gel picture regarding protein
>
> Sivasankar Putta
>
--
Christian Roth
Institut für Bioanalytische Chemie
Biotechnologisch-Biomedizinisches Zentrum
Fakultät für Chemie und Mineralogie
Universität Leipzig
Deutscher Platz 5
04103 Leipzig
Telefon: +49 (0)341 97 31316
Fax: +49 (0)341 97 31319
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