Protein degradation in E.coli

[John Lum]

Hi there,

> Do you know when the degradation is happening? I think degradation is
> quite common for GST fusions. For some fusions I made, the degradation
> only happened during cell lysis- as seen in crude soluble extracts, but
> not in total protein from boiled cells. Try different extraction
> conditions if this is the case. I found degradation to be much reduced
> when 1 mM DTT was included and Triton-X100 left out of the lysis buffer.

       I'm analysing the total protein from boiled cells. When I use the GST
antibody for western, I found multiple bands of smaller size.

Regards,
John