Any protein will absorb at 230, and most UV detectors these days can
reach that, if you want to follow that with UV.
Together with SDS_PAGE of course, this should be good enough to go
ahead.
A.
On May 28, 2010, at 18:03, 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, Ile
(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
