I want to check whether my protein is undergoing in vitro phosphorylation by substrate which is a Kinase. I am working on plant proteins. I am planning to do the reaction as follows,

MBP-Protein (X)

GST-Kinase (Substrate)

ATP

1X PBS

Reaction at 30C for 1 hr. detection by Anti-MBP.

Can anyone who has experience of doing In vitro Phosphorylation of Plant proteins kindly share with me?

Note: I checked the phosphorylaiton site by in silico (NetPhos 2.0).

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