Revision 1
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:

#Q13535

Entrez-Gene Id:

545

Product Includes Quantity Reactivity MW(kDa) Isotype
ATR (E1S3S) Rabbit mAb 13934 100 µl H M R 300 Rabbit IgG
Phospho-ATR (Thr1989) (D5K8W) Rabbit mAb 30632 100 µl H 300 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 antibody.

Background

Ataxia telangiectasia mutated kinase (ATM) and ataxia telangiectasia and Rad3-related kinase (ATR) are PI3 kinase-related kinase (PIKK) family members that phosphorylate multiple substrates on serine or threonine residues that are followed by a glutamine in response to DNA damage or replication blocks (1-3). Despite the essential role of ATR in cell cycle signaling and DNA repair processes, little is known about its activation. ATR was long thought to exist in a constitutively active state in cells, with DNA damage-induced signaling occurring via recruitment of ATR to single stranded DNA and sites of replication stress. Phosphorylation of ATR at serine 428 in response to UV-induced DNA damage has been suggested as a means of activating ATR (4,5). Recent work has shown autophosphorylation of ATR at threonine 1989. Like ATM Ser1981, phosphorylation of ATR Thr1989 occurs in response to DNA damage, indicating that phosphorylation at this site is important in ATR-mediated signaling (6,7).

  1. Kastan, M.B. and Lim, D.S. (2000) Nat Rev Mol Cell Biol 1, 179-86.
  2. Abraham, R.T. (2004) DNA Repair (Amst) 3, 883-7.
  3. Shechter, D. et al. (2004) DNA Repair (Amst) 3, 901-8.
  4. Vauzour, D. et al. (2007) Arch Biochem Biophys 468, 159-66.
  5. Smith, J. et al. (2010) Adv Cancer Res 108, 73-112.
  6. Nam, E.A. et al. (2011) J Biol Chem 286, 28707-14.
  7. Liu, S. et al. (2011) Mol Cell 43, 192-202.

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.
    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 1
    #53217

    PhosphoPlus® ATR (Thr1989) Antibody Duet

    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from HeLa + Hydroxyurea (1.5mM, 17hr) cells using Phospho-ATR (Thr1989) (D5K8W) Rabbit mAb #30632. The virtual lane view (left) shows a single target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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 66-440 kDa separation module.
    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 2 Expand Image
    Western blot analysis of extracts from various cell lines using ATR (E1S3S) Rabbit mAb.
    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 3 Expand Image
    Western blot analysis of extracts from HeLa cells, untreated (-) or treated with hydroxyurea (1.5 mm, 16 hr; +), using Phospho-ATR (Thr1989) (D5K8W) Rabbit mAb (upper) or ATR (E1S3S) Rabbit mAb #13934 (lower).
    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 4 Expand Image
    Immunoprecipitation of ATR from HT-29 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or ATR (E1S3S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using ATR Antibody #2790.