Is your protein in the membrane?. I did in the past western blots trying
to detect GPCR tagged with Flag, HA or His epitopes in expression
systems such as Cos or 293 and I got nothing. By that time I thought
either the protein was degraded during membrane isolation, was not
completely solubilized or the antibodies were not strong enought to
resist the washing steps I was using because the receptor was expressed
in binding assays (>1 pmol/mg prot)
Greatings
Marisol
--
MS Rodriguez Pena, MD, phD
Department of Pharmacochemistry, Faculteit der Scheikunde
Vrije Universiteit.
De Boelelaan 1083,
1081 HV Amsterdam, The Netherlands
Tel: 31-20-4447572
Fax: 31-20-4447610
