Is a tagged protein more prone to degradation? (Lysis protocols??)

Marisol Rodriguez Pena pena at chem.vu.nl
Wed Nov 19 08:04:00 EST 1997


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



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