A versatile microtiter assay for the universal cdc2 cell cycle regulator.
A microassay for p34cdc2 based on the high affinity association between cdc2 and Schizosaccharomyces pombe p13suc1 has been developed. p13 purified from Escherichia coli was immobilized on microtiter plates and cellular lysate was incubated in the wells to allow the binding of cdc2 and its associated proteins. p34cdc2 was assayed either as a histone kinase or by immunological methods. The method was optimized for S. pombe cell extracts but can also be applied to other organisms such as Xenopus oocytes or HeLa cells. This rapid assay allows the specific determination of p34cdc2 histone H1 kinase activity in a very large number of samples.
Ducommun
B
B
Cold Spring Harbor Laboratory, New York 11724.
Beach
D
D
D000818:Animals; D016203:CDC2 Protein Kinase; D002453:Cell Cycle / Q000502:physiology*; D018797:Cell Cycle Proteins; D002460:Cell Line; D005656:Fungal Proteins; D006367:HeLa Cells; D006801:Humans; D007124:Immunoenzyme Techniques; D009865:Oocytes; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism; D006656:Protamine Kinase / Q000378:metabolism; D012440:Saccharomyces; D029702:Schizosaccharomyces pombe Proteins; D014981:Xenopus
D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D010750:Phosphoproteins; D029702:Schizosaccharomyces pombe Proteins; C051655:Suc1 protein, S pombe; D006656:Protamine Kinase; D016203:CDC2 Protein Kinase