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
