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Open Forum on Methodology

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Antibodies to pMAPK

15 December 2000

Eve De Lamirande

I am also very interested in these bands that are recognized by the andibody raised against the doubly phosphorylated TEY.

I have problems to believe that this is non specific binding. There are many bands labeled and the changes in intensity that I observe appear too much related to activation of signal transduction to be non specific binding. I did a time course study and it confirmed my idea that it cannot be only non specific binding.

I also tried few conditions to do the immunoblotting and I get almost always the same thing.

What puzzles me the most is that this type of TEY phosphorylation is performed by dual specificity kinase such as MEK and that ERK1/2 is supposed to be the only "known" substrate for MER (this is what I read ... is it true?). When I add PD98059 to the cells, there is an effect on one protein band labeled by anti-*TEY*, but it is not one of ERK1/2.

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