Hi,
I would appreciate any help on the following topic:
I am trying to run several neuropeptides on SDS-PAGE: neuropeptide Y
(Peninsula), vasoactive intestinal polypeptide (Peninsula), substance
P (Sigma) and Met-enkephalin (Sigma). These are not tissue extracts
but just pure peptides.
I tried different recipes for gels, and now I used method described by
Schaegger and von Jagow (Anal Biochem, 166:368-379, 1987) - 16.5%T3%C
separating, 10T3C spacer and 4T3C stacking gels. Schaegger and von
Jagow claim it has good resolution from 1kDa to 70kDa. After
electrophoresis I stain gel in regular CBB R-450 (with methanol). Now
strange thing comes: I cannot see a band of Met-enkephalin (maybe
nothing strange - just 0.6 kDa), I cannot see band of substance P
(just ca. 1.3 kDa) !!!, AND bands of neuropeptide Y (4.25kDa) and
vasoactive intestinal polypeptide (3.3 kDa) run at EXACTLY the same
position.
Does anyone have some experience with separation of thus small
peptides?
ANY help and suggestion will be appreciated.
Krzysztof W. Wasowicz
Department of Animal Anatomy
University of Warmia & Mazury
ul. Oczapowskiego 13 (Bldg. 105J)
10-957 Olsztyn-Kortowo
Poland
Phone: +48-89-5233688
Fax: +48-89-5234986
e-mail: wasowicz at moskit.uwm.edu.pl
ICQ: 58434323
