Hi Yogi,
You can see your peptide on a gel so why can't you "monitor" it by SDS-PAGE? A little time consuming, yes, but then you have the extra benefit of also seeing if there are contaminating proteins in your sample.
good luck,
Eric
__________________________
Eric Larson, PhD
MSGPP Consortium
Department of Biochemistry
Box 357742
University of Washington
Seattle, WA 98195
On Fri, 28 May 2010, Sollepura Yogesha wrote:
>
> Dear All,
>
> I have expressed 30-40 aa region my protein fused to GST.
>
> I subjected it to precision protease cleavage. On the gel I can see the band.
>
> When I looked for ProtParam in expasy it shows that my peptide doesn’t have Extinction coefficients as “ there are no Trp, Tyr or
> Cys in the region considered, your protein should not be visible by UV spectrophotometry.”
>
> I need to separate GST from the cleavage mixture.
>
> How can I monitor my peptide during FPLC and after that.
>
> AA composition is Ala (A) 8, Arg (R) 2, Asn (N) 3, Asp (D) 2, Gln (Q) 2, Glu (E) 2, Gly (G) 4, I
> le (I) 1, Leu (L) 4, Lys (K) 4, Phe (F) 2, Pro (P) 2, Ser (S) 5, Thr (T) 5, Val (V) 3.
>
> I am looking for some suggestions
>
> Thanks in advance
>
> Yogi
>
>
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