Assaying Cdc25 phosphatase activity.
To determine the activity of all human Cdc25 phosphatases, two different methods are described. For assaying phosphatase activities of recombinant Cdc25 proteins produced in Escherichia coli or insect cells, a fluorimetric assay using fluorescein diphosphate (FDP) as a substrate is recommended. To analyze endogenous Cdc25 phosphatase activities of immunoprecipitates from total cellular extracts, the physiological substrate Cdk1/cyclin B1 is most sensitive.
Hassepass
Ingo
I
Cell Cycle Control and Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg.
Hoffmann
Ingrid
I
D000818:Animals; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D019926:Cyclin B / Q000378:metabolism; D056744:Cyclin B1; D004926:Escherichia coli / Q000378:metabolism; D019793:Fluorescein / Q000378:metabolism; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D016196:S Phase; D018411:Spodoptera / Q000166:cytology / Q000235:genetics; D020687:cdc25 Phosphatases / Q000032:analysis*
C534699:CCNB1 protein, human; D019926:Cyclin B; D056744:Cyclin B1; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D011494:Protein Kinases; C102333:histone H1 kinase; D016203:CDC2 Protein Kinase; D020687:cdc25 Phosphatases; D019793:Fluorescein