Immuno-precipitation of Phospho-Tyr proteins

I am trying to assess the degree of phosphorylation a receptor
undergoes in response to it's ligand plus various other factors.
If I apply the ligand to my cells and then extract in the presence of
sodium orthovanadate and do a western using an antibody against
phospho-tyrosine I see a band appearing which corresponds to the size
of my receptor and which doesn't appear in the non ligand- treated cell extract. However if I immunoprecipitate the receptor (in the presence of orthovanadate) I appear to lose the signal with the phospho-tyrosine antibody although there is clearly receptor present in the treated and untreated ... ...

Last update 21-Nov-2000, Rating of 0 votes.

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