Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Antibody Pair #7838

Kit Includes Volume Cap Color
cdc2 (Tyr 15) Rabbit Capture Antibody (100X) 0.4 milliliters Pink
cdc2 Mouse Detection Antibody (100X) 0.4 milliliters Blue
Anti-Mouse IgG, HRP-Linked Antibody (1000X) 0.04 milliliters Yellow

Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human

Description

CST's PathScan® Phospho-cdc2 (Tyr15) Sandwich ELISA Antibody Pair is being 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 this kit are custom formulations specific to the kit.

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.

Background

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 Tyr15 and Thr14 (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).

  1. Atherton-Fessler, S. et al. (1994) Mol. Biol. Cell. 5, 989-1001.
  2. Norbury, C. et al. (1991) EMBO. J. 10, 3321-3329.
  3. McGowan, C.H. and Russell, P. (1993) EMBO J. 12, 75-85.
  4. Wells, N.J. et al. (1999) J. Cell. Sci. 112, 3361-3371.
  5. Hunter, T. (1995) Cell 80, 225-236.

Application References

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This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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