Preservation of protein phosphoryl groups in immunoprecipitation assays.
A phosphorylation-sensitive immunoprecipitation protocol is presented which ensures quantitative and qualitative recovery of phosphoantigens from different phosphatase environments. The protocol is based on the phosphatase-inactivating properties of a specific mixture of chemical compounds included in the "classical" immunoprecipitation buffers.
Papavassiliou
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AG
Differentiation Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
D002478:Cells, Cultured; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D010749:Phosphoprotein Phosphatases / Q000737:chemistry*; D010766:Phosphorylation; D011233:Precipitin Tests / Q000379:methods*; D016760:Proto-Oncogene Proteins c-fos / Q000032:analysis; D016755:Proto-Oncogene Proteins c-jun / Q000032:analysis; D014162:Transfection
D016760:Proto-Oncogene Proteins c-fos; D016755:Proto-Oncogene Proteins c-jun; D010749:Phosphoprotein Phosphatases