Words similar to prb-directed
Example sentences for: prb-directed
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Preparation of [ 32P]pRB substrate and pRB-directed phosphatase activity assays
Under conditions favoring proliferation, the phosphorylation of pRB via cyclin-dependent kinases (CDKs) in mid-to-late G 1 phase results in liberation of E2F and other pRB-bound transcription factors, which then activate the transcription of S-phase genes [ 19 ] . In late M phase, pRB returns to its growth suppressive, hypophosphorylated form due to the action of a pRB-directed protein phosphatase belonging to the type 1 class of serine/threonine protein phosphatases [ 11 20 21 ] .
To assay pRB-directed phosphatase activity, [ 32P]-labeled pRB substrate was prepared by immunoprecipitation with monoclonal antibody to pRB, PMG3-245 (PharMingen), from [ 32P]-labeled CV-1P cells as described previously [ 20 ] . For the source of enzyme, immunocomplexed 6His-HA-PP1α from induced LLWO2F cells were mixed and incubated with [ 32P]-labeled immunoprecipitated pRB for 30 minutes at 30°C as previously described [ 20 ] . Phosphatase reactions were terminated by the addition of SDS-PAGE sample buffer and boiling.