title: Measurement of Cdk1/Cyclin B Kinase Activity by Specific Antibodies and Western Blotting.
abstract: Quantitative measurement of enzyme activity is a valuable approach to study how cells function. We present a method to measure the activity of the enzyme Cdk1/cyclin B. This enzyme is required by all eukaryotic cells to enter mitosis. Therefore, a biochemical assay to measure Cdk1/cyclin B activity can be used to identify cell populations that are in mitosis or to detect inhibitors of Cdk1/Cyclin B in vitro. A key distinction of the method presented here, compared to others, is that it uses a recombinant protein, a specific antibody, and a western blot apparatus, which makes the technique available to cell and molecular biology laboratories who do not wish to use radioisotopes, which are commonly required for other protein kinase assays.
authors.affiliation: Cancer Cell Laboratory, Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, AB, Canada, T1K 3M4.
authors.forename: Cody W
authors.initials: CW
authors.lastname: Lewis
authors.forename: Ryan G
authors.initials: RG
authors.lastname: Taylor
authors.forename: Roy M
authors.initials: RM
authors.lastname: Golsteyn
mesh_terms: D000918:Antibody Specificity; D015153:Blotting, Western / Q000379:methods*; D016203:CDC2 Protein Kinase / Q000037:antagonists & inhibitors / Q000276:immunology* / Q000378:metabolism*; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D019926:Cyclin B / Q000276:immunology* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays / Q000379:methods*; D006801:Humans; D008938:Mitosis / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology
chemical_list: D019926:Cyclin B; D047428:Protein Kinase Inhibitors; D016203:CDC2 Protein Kinase