kinases PY-blot

[marianne.fjording@leo-pharma.com]

Hello

Does anybody have a solution for reducing the background on a phosphotyrosine
western blot?

We have i.p. with an anti-VEGF-R antibody (Santa Cruz) and blot with monoclonal
4G10 (UBI) phosphotyrosine antibody. We have also tried with PY-20
(Transduction).

We have tried to block in 5% skimmilk or 3% BSA. We do the washing with PBS+0.1%
Tween-20, and develop with ECL.

Also, is there anybody doing a VEGF-R kinase assay using phosphotyrosine
antibodies or phosphorylation of a substrate such as Poly-Glu-Tyr?

Thank you
Marianne Scheel Fjording
LEO Pharmaceutical Products
Denmark