{"product_id":"revvity-64cdk2y15peh","title":"Revvity, 64CDK2Y15PEH, HTRF Human and Mouse Phospho-CDK2 (Tyr15) Detection Kit, 10,000 Assay Points","description":"\u003cp\u003eOverview\u003cbr\u003e\nThis HTRF cell-based assay conveniently and accurately detects phosphorylated CDK2 at Tyr15. ​​​​​​​\u003cbr\u003e\nCDK2 (Cyclin-Dependent Kinase 2) is a member of the subfamily of CDKs that coordinate cell cycle progression in mammalian cells (also including CDK1, CDK4, and CDK6). CDK2 is activated by interaction with Cyclin E and Cyclin A during the late G1 phase and S phase respectively, and is also regulated by two major phospho-sites. Phosphorylation at Tyr15 by the kinase Wee1 is inhibitory by preventing ATP binding, whereas Tyr15 dephosphorylation by the phosphatase Cdc25A and phosphorylation at Thr160 by CAK (CDK Activating Kinase) is required for its full activity.\u003cbr\u003e\nCDK2 inhibitory phosphorylation at Tyr15 is essential for maintaining genome integrity and preventing DNA damage during the S phase. The Wee1\/Cdc25A axis is therefore an attractive target for cancer therapy and may represent a unique approach to sensitize cancer cells with hyperactive CDK2.\u003cbr\u003e\nHow it works\u003cbr\u003e\nPhospho-CDK2 (Tyr15) assay principle\u003cbr\u003e\nThe Phospho-CDK2 (Tyr15) assay measures CDK2 when phosphorylated at Tyr15. Unlike Western Blot, the assay is entirely plate-based and does not require gels, electrophoresis, or transfer. The assay uses 2 antibodies, one labeled with a donor fluorophore and the other with an acceptor. The first antibody was selected for its specific binding to the phosphorylated motif on the protein, and the second for its ability to recognize the protein independently of its phosphorylation state. Protein phosphorylation enables an immune-complex formation involving both labeled antibodies, and which brings the donor fluorophore into close proximity to the acceptor, thereby generating a FRET signal. Its intensity is directly proportional to the concentration of phosphorylated protein present in the sample, and provides a means of assessing the protein's phosphorylation state under a no-wash assay format.\u003c\/p\u003e\n\n\n\n\n  \u003cbr\u003e\n        \n\n\n\t\u003cbr\u003e\n  \u003cbr\u003e\n        \u003cbr\u003e\n  \n\n      \u003cbr\u003e\n  \u003cbr\u003e\n  \u003cbr\u003e\n    \u003cbr\u003e\n      \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n      \u003cbr\u003e\n    \n\n    \u003cbr\u003e\n      \n\n      \u003cbr\u003e\n        \n\n      \u003cbr\u003e\n    \n\n  \n\n\n\n\n\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003ePhospho-CDK2 (Tyr15) two-plate assay protocol\u003cbr\u003e\nThe two-plate protocol involves culturing cells in a 96-well plate before lysis, then transferring lysates into a 384-well low volume detection plate before the addition of Phospho-CDK2 (Tyr15) HTRF detection reagents. This protocol enables the cells' viability and confluence to be monitored.\u003c\/p\u003e\n\n\n\n\n  \u003cbr\u003e\n        \n\n\n\t\u003cbr\u003e\n  \u003cbr\u003e\n        \u003cbr\u003e\n  \n\n      \u003cbr\u003e\n  \u003cbr\u003e\n  \u003cbr\u003e\n    \u003cbr\u003e\n      \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n        \u003cbr\u003e\n      \u003cbr\u003e\n    \n\n    \u003cbr\u003e\n      \n\n      \u003cbr\u003e\n        \n\n      \u003cbr\u003e\n    \n\n  \n\n\n\n\n\u003cp\u003e \u003c\/p\u003e\n\n\n\n\n\u003cp\u003ePhospho-CDK2 (Tyr15) one-plate assay protocol\u003cbr\u003e\nDetection of Phosphorylated CDK2 (Tyr15) with HTRF reagents can be performed in a single plate used for culturing, stimulation, and lysis. No washing steps are required. This HTS designed protocol enables miniaturization while maintaining robust HTRF quality. Application-Cell Signaling\u003cbr\u003e\nBrand-HTRF\u003cbr\u003e\nDetection Modality-HTRF\u003cbr\u003e\nLysis Buffer Compatibility-Lysis Buffer 2 Lysis Buffer 3 Lysis Buffer 4 Lysis Buffer 5\u003cbr\u003e\nMolecular Modification-Phosphorylation\u003cbr\u003e\nProduct Group-Kit\u003cbr\u003e\nSample Volume-16 µL\u003cbr\u003e\nShipping Conditions-Shipped in Dry Ice\u003cbr\u003e\nTarget Class-Phosphoproteins\u003cbr\u003e\nTarget Species-Human Mouse\u003cbr\u003e\nTechnology-TR-FRET\u003cbr\u003e\nTherapeutic Area-Oncology \u0026amp; Inflammation\u003cbr\u003e\nUnit Size-10,000 assay points\u003c\/p\u003e","brand":"Revvity","offers":[{"title":"Default Title","offer_id":46775240982697,"sku":"64CDK2Y15PEH","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/revvity-64cdk2y15peh","provider":"Iright","version":"1.0","type":"link"}