The relationship between protein concentration of lysates from λ phosphatase-treated and IL-4-treated HeLa cells and the absorbance at 450 nm using PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Antibody Pair #7838 is shown. HeLa cells (80% confluent) were treated with human IL-4 for 10 minutes (100 ng/ml) at 37°C and then lysed.Learn more about how we get our images
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
1X Cell Lysis Buffer: 10X Cell Lysis Buffer (#9803): To prepare 10 ml of 1X Cell Lysis Buffer, add 1 ml of 10X Cell Lysis Buffer to 9 ml of dH2O, mix. Buffer can be stored at 4°C for short-term use (1–2 weeks).
Recommended: Add 1 mM phenylmethylsulfonyl fluoride (PMSF) (#8553) immediately before use.
STOP Solution: (#7002)
NOTE: Reagents should be made fresh daily.
posted January 2008
revised Sepetember 2013
|Product Includes||Volume||Cap Color|
|cdc2 (Y15) Capture Ab (100X)||400 µl||Pink|
|cdc2 Detection Ab (100X)||400 µl||Blue|
|Anti-mouse IgG, HRP-linked Antibody (1000X)||40 µl||Yellow|
CST's PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Kit #7176. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-cdc2 (Tyr15) Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a cdc2 Detection Antibody and anti-Mouse IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho-cdc2 (Tyr15) protein.
Antibodies in kit are custom formulations specific to kit.
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The entry of eukaryotic cells into mitosis is regulated by cdc2 kinase activation, a process controlled at several steps including cyclin binding and phosphorylation of cdc2 at Thr161 (1). However, the critical regulatory step in activating cdc2 during progression into mitosis appears to be dephosphorylation of cdc2 at Thr14 and Tyr15 (2). Phosphorylation at Thr14 and Tyr15, resulting in inhibition of cdc2, can be carried out by Wee1 and Myt1 protein kinases (3,4). The cdc25 phosphatase may be responsible for removal of phosphates at Thr14 and Tyr15 and subsequent activation of cdc2 (1,5).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. PathScan is a trademark of Cell Signaling Technology, Inc. U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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