Anti-phosphotyrosine immunoblot

Jim Hutchins hutchins at netdoor.com
Fri Aug 23 20:59:17 EST 1996


Cornelius Krasel (krasel at wpxx02.toxi.uni-wuerzburg.de) wrote:
: John Hines (hines at pharm.med.upenn.edu) wrote:
[regarding problems with anti-PY antibodies in Western blotting]
: > Am I the only one with such problems with ECL??
: 
: No. Occasionally ECL gives strange blots (i.e. plenty of background). 
: This may depend on your blocking conditions (we use 2% milk 1% ovalbumin
: and incubate our 1st antibody usually in this solution; you may also
: try to increase blocking time, e.g. overnight). 

Just a slight (but important) correction: milk contains phosphotyrosine,
so block in 2% BSA or similar, *NOT* milk.

-- 
Jim Hutchins    hutchins at netdoor.com   hutchins at umsmed.edu
http://fiona.umsmed.edu/~hutchins/



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