METHODS FOR MEASURING CDC25B PHOSPHATASE ACTIVITY AND METHODS FOR SCREENING CDC25B PHOSPHATASE INHIBITORS USING THE SAME

摘要

After the present invention by reacting the entire immune saliva of CyclinB combined with the protein extract and Cdk1 obtained by culturing the HeLa cell line expressing the Cdc25B2 or Cdc25B3 (immunoprecipitant) leads to dephosphorylation of the 15th tyrosine residue of Cdk1, non-radiation measurement performing the method of Western blotting to provide a method measuring the activity of Cdc25B phosphatase comprising the step of measuring the activity of Cdk1 kinase and phosphatase inhibitors Cdc25B search method using the same. In addition, the present invention then by reacting the protein extract and the whole immune saliva of Cdk1 obtained by culturing the HeLa cell line expressing the Cdc25B2 or Cdc25B3 induce dephosphorylation of Cdk1, to measure the 15 de-phosphorylation level of tyrosine residues of Cdk1 method activity measurement of Cdc25B phosphatase, comprising the step, and provides a choice of Cdc25B phosphatase inhibitors method using the same.; The test method and the search method of the present invention is the specificity caused by the lack of post-translational human Cdc25B from the use of Cdc25B inappropriate reflection and recombinant fusion activity Cdc25B protein due to non-specific substrate used in the conventional formulation (Post-translational modification) developing a can overcome the degradation.