title: Assaying Cdc25 phosphatase activity.
abstract: 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.
authors.affiliation: Cell Cycle Control and Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg.
authors.forename: Ingo
authors.initials: I
authors.lastname: Hassepass
authors.forename: Ingrid
authors.initials: I
authors.lastname: Hoffmann
mesh_terms: 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*
chemical_list: 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