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.

    限制使用

    除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

    专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

    Revision 1
    #53217

    PhosphoPlus® ATR (Thr1989) Antibody Duet

    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 1 Expand Image
    使用 Phospho-ATR (Thr1989) (D5K8W) Rabbit mAb #30632 对来自 HeLa + Hydroxyurea(1.5mM,17 小时)细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:50 和 1:250 时的单一靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:50(蓝线)和 1:250(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 66-440 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 2 Expand Image
    使用 ATR (E1S3S) Rabbit mAb 对不同细胞系提取物进行蛋白质印迹分析。
    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 3 Expand Image
    使用 Phospho-ATR (Thr1989) (D5K8W) 兔单克隆抗体(上图)或 ATR (E1S3S) 兔单克隆抗体 #13934(下图)对未经 (-) 或已经羟基脲(1.5 mm,16 小时;+)处理的 HeLa 细胞的提取物进行蛋白印迹分析。
    PhosphoPlus® ATR (Thr1989) Antibody Duet: Image 4 Expand Image
    使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (泳道 2) 或 ATR (E1S3S) Rabbit mAb (泳道 3) 对 HT-29 细胞提取物的 ATR 进行免疫沉淀 。泳道 1 是 10% 输入对照。使用 ATR Antibody #2790 进行蛋白质印迹分析。