Revision 2
Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
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UniProt ID:

#O14757

Entrez-Gene Id:

1111

Product Includes Quantity Reactivity MW(kDa) Isotype
Chk1 (2G1D5) Mouse mAb 2360 100 µl H M R Mk 56 Mouse IgG1
Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb 12302 100 µl H M Mk 56 Rabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications. 

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibodies.

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 by ATM/ATR, followed by autophosphorylation of Ser296. Activation 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.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    PhosphoPlus is a registered trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 2
    #51664

    PhosphoPlus® Chk1 (Ser317) Antibody Duet

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 1 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from HeLa cells using Chk1 (2G1D5) Mouse mAb #2360. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 2 Expand Image
    Western blot analysis of extracts from 293 and NIH/3T3 cells, untreated (-) or UV-treated (100 mJ, 1 hr recovery; +), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb. The blot on the right was treated with calf intestinal phosphatase (CIP) before western blot.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using Chk1 (2G1D5) Mouse mAb.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 4 Expand Image

    Immunohistochemical analysis of paraffin-embedded human oropharyngeal squamous cell carcinoma using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 5 Expand Image
    Immunoprecipitation of phospho-Chk1 (Ser317) from 293 cell extracts treated with UV (100 mJ, 1 hr recovery) using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (lane 2) or Rabbit (D1AG) mAb IgG XP® Isotype Control #3900 (lane 3). Lane 1 is 10% input.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 6 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Chk1 siRNA I #6241 or SignalSilence® Chk1 siRNA II (+), using Chk1 (2G1D5) Mouse mAb #2360 and β-Actin (13E5) Rabbit mAb #4970. Chk1 (2G1D5) Mouse mAb confirms silencing of Chk1 expression and β-Actin (13E5) Rabbit mAb is used to control for loading and specificity of Chk1 siRNA.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 7 Expand Image

    Immunohistochemical analysis of paraffin-embedded human thymoma using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 8 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left), UV-treated (center), or UV and λ phosphatase-treated (right), using Phospho-Chk1 (Ser317) (D12H3) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 9 Expand Image
    Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Chk1 (2G1D5) Mouse mAb.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 10 Expand Image

    Immunohistochemical analysis of paraffin-embedded human esophageal adenocarcinoma using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 11 Expand Image

    Immunohistochemical analysis of paraffin-embedded human Non-Hodgkin lymphoma using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 12 Expand Image

    Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 13 Expand Image

    Immunohistochemical analysis of paraffin-embedded human gastric adenocarcinoma using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 14 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrial cancer using Chk1 (2G1D5) Mouse mAb.
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 15 Expand Image

    Immunohistochemical analysis of paraffin-embedded normal human uterus using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 16 Expand Image

    Immunohistochemical analysis of paraffin-embedded human tonsil using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 17 Expand Image

    Immunohistochemical analysis of paraffin-embedded normal human esophagus using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 18 Expand Image

    Immunohistochemical analysis of paraffin-embedded normal human testis using Chk1 (2G1D5) Mouse mAb.

    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 19 Expand Image
    Immunohistochemical analysis of paraffin-embedded normal human small intestine using Chk1 (2G1D5) Mouse mAb (left) compared to concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (right). 
    PhosphoPlus® Chk1 (Ser317) Antibody Duet: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded 293T cell pellet (left, high-expressing) or SK-MEL-28 cell pellet (right, low-expressing) using Chk1 (2G1D5) Mouse mAb.