How to prevent proteolytic degradation

Alejandro Miguel Martin Dunn alejandro.martin at cigb.edu.cu
Thu Nov 9 22:10:43 EST 2000


>
>Choe Woo-seok wrote:
(...)
>> When the IB protein is dissolved in 50 mM Tris, 50 mM DTT, 
>8M Urea, pH 8.0,
>> the proteolytic
>> degradation of target protein (Mw = 65 kd, with 6xHis tag) 
>started.  We
(...)
>> Is there anybody who has experienced the similar problem and 
>sorted it out
>> finally?
I remember a colleague of mine having exactly that problem (the protein
starts degrading as soon as it is solubilized in urea). As far as my
memory goes, she solved the problem using a different chaotrope (I think
it was GuHCl).
I'll ask her tomorrow and let you know if anything new comes out.

cheers,
Alejandro

---------------------------------------------
Alejandro Martin
Centro de Ingenieria Genetica y Biotecnologia
PO Box 6162, C. Postal 10600
Ciudad Habana, CUBA
Tel: (53-7)216221, FAX: (53-7)214764

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