cdc2 (Tyr15) BioAssay(TM) Phosphorylation Detection Kit

Entry of all eukaryotic cells into M-phase of the cell cycle is regulated by activation of cdc2 kinase (figure 1). Activation of cdc2 is controlled at several steps including cyclin binding and phosphorylation of threonine 161 (1-4). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of Tyr15 and Thr14 (3,5). Phosphorylation at Tyr15 and inhibition of cdc2 is carried out by WEE1 and MIK protein kinases while Tyr15 dephosphorylation and activation of cdc2 is carried out by the cdc25 phosphatase (3,4,6).\n\nApplications: \nSuitable for use in Western Blotting. Other applications not tested.\n\nRecommended Dilution:\nWestern blotting: 1:1000\n\nOptimal dilutions to be determined by the researcher.\n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage, store at -20 degrees C. Aliquots are stable for at least 6 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

C2549-55

Size

1Kit

Applications

WB

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, BSA, 50% glycerol.

Purity

Purified by protein A and peptide affinity chromatography.

References

1. Norbury, C., and Nurse, P. (1992) Annu. Rev. Biochem. 61, 441

Additional Info

When used in conjunction with the HRP Western Detection System, phospho-cdc2 (Tyr15) antibody detects less than 10ng of tyrosine phosphorylated cdc2 yet will not react with up to 1ug of non-tyrosine phosphorylated cdc2. Similarly, phospho-cdc2 (Tyr15) antibody detects phosphorylated tyrosine 15 of cdk2 but demonstrates no crossreactivity to cdk4, cdk6 and cdk7 or other phospho-tyrosine proteins.