desperately need help
KeldS at uic.edu
Mon Mar 10 08:26:07 EST 1997
> Hi people who are expert for western!
> I am doing western using PY20 (antiphosphotyrosine antibody) and
> developing with ECL but it did not work. I checked every condition it is
> still not working.
> Any suggestion is appreciated.
> Thank you
Did you use milk as the blocking reagent? Milk has phosphoproteins and
could give you a problem. You have to give specifics though - right now
can help you - it would be like bringing your car in for repair and say:
"My car is not working right, please fix it!" -
So please explain the details of your WB along with observations
- details such as:
how much sample
transferred to what membrane
what conc. of PY20
what enz-ab conjugate or are you suing PY-20 enzyme conjugate
What substrate - etc etc etc.
More information about the Methods