Cdc25B phosphatase activity measurements and search for ways of Cdc25B phosphatase inhibitors using the same

摘要

The present invention is combined with a protein extract and Cdk1 obtained by culturing the HeLa cell line expressing Cdc25B2 or Cdc25B3 reacting the immune whole saliva (immunoprecipitant) of CyclinB induced the de-phosphorylation of tyrosine residues in the 15th and Cdk1, by performing the measurement method of the non-radiation-Western blotting of Cdc25B phosphatase comprising the step of measuring the activity of kinases Cdk1 provides a search method of the activity measurement method using the same, and Cdc25B phosphatase inhibitors. In addition, the present invention for measuring the degree of dephosphorylation of the tyrosine residues 15th Cdc25B2 or cultured HeLa cell line expressing Cdc25B3 by reacting the immune saliva and whole of the resulting protein extract induced the dephosphorylation of Cdk1 and Cdk1, Cdk1 step provides a search method in Cdc25B phosphatase inhibitor activity measured using this method and the Cdc25B phosphatase comprising a. ; The test method and the search method of the present invention is due to non-specific activity of Cdc25B substrate used in the conventional After the translation of human reflection and Cdc25B caused by improper use of the recombinant fusion protein Cdc25B can overcome the degradation caused by the lack of specificity of the formulation (Post-translational modification) symptoms.