Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total Chk1 Sandwich ELISA Antibody Pair #7873

Kit Includes Volume Cap Color
Chk1 Capture Ab (100X) 0.4 ml Pink
Chk1 Detection Ab (100X) 0.4 ml Blue
Anti-rabbit IgG, HRP-linked Antibody (1000X) 0.04 ml Red

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H

Reactivity Key:  H=Human
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Important Ordering Details

Product is assembled upon order to ensure maximum activity. Domestic: Please allow up to two weeks for your order to be processed and shipped. International: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.

Protocols

Description

CST's PathScan® Total Chk1 Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Total Chk1 Sandwich ELISA Kit #7872. Capture and detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Chk1 mouse capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysates are added, followed by Chk1 detection antibody and anti-rabbit IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of total Chk1.Antibodies in kit are custom formulations specific to kit.

Specificity / Sensitivity

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.

Sandwich ELISA

Sandwich ELISA

The relationship between lysate protein concentration from untreated and UV treated HeLa cells and the absorbance at 450 nm using PathScan® Total Chk1 Sandwich ELISA Antibody Pair #7873 is shown. HeLa cells were treated with UV at 100 mj, incubated for 60 minutes at 37ºC and then lysed.

Background

Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 and occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for re-entry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of aurora B and BubR1 (8). Research studies have implicated Chk1 as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9).

  1. Liu, Q. et al. (2000) Genes Dev 14, 1448-59.
  2. Zhao, H. and Piwnica-Worms, H. (2001) Mol Cell Biol 21, 4129-39.
  3. Jiang, K. et al. (2003) J Biol Chem 278, 25207-17.
  4. Martin, S.A. and Ouchi, T. (2008) Mol Cancer Ther 7, 2509-16.
  5. Chen, M.S. et al. (2003) Mol Cell Biol 23, 7488-97.
  6. Zeng, Y. et al. (1998) Nature 395, 507-10.
  7. Löffler, H. et al. (2006) Cell Cycle 5, 2543-7.
  8. Zachos, G. et al. (2007) Dev Cell 12, 247-60.
  9. Garber, K. (2005) J Natl Cancer Inst 97, 1026-8.

Application References

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Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

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