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.
Applications:

WB, IP, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

65, 75

Source/Isotype:

Rabbit IgG

UniProt ID:

#P17544, #P15336

Entrez-Gene Id:

11016, 1386

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:100
Immunofluorescence (Immunocytochemistry) 1:800 - 1:3200
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800

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.

Specificity / Sensitivity

Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb detects endogenous levels of ATF-2 and ATF-7 only when phosphorylated at Thr69 and Thr51, respectively. It recognizes Thr69/Thr71 dually phosphorylated ATF-2, Thr51/Thr53 dually phosphorylated ATF-7, Thr69 singly phosphorylated ATF-2, and Thr51 singly phosphorylated ATF-7 equally well but does not detect singly phosphorylated Thr71 of ATF-2 and singly phosphorylated Thr53 of ATF-7.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr69 and Thr71 of human ATF-2 protein.

Background

The transcription factor ATF-2 (also called CRE-BP1) binds to both AP-1 and CRE DNA response elements and is a member of the ATF/CREB family of leucine zipper proteins (1). ATF-2 interacts with a variety of viral oncoproteins and cellular tumor suppressors and is a target of the SAPK/JNK and p38 MAP kinase signaling pathways (2-4). Various forms of cellular stress, including genotoxic agents, inflammatory cytokines, and UV irradiation, stimulate the transcriptional activity of ATF-2. Cellular stress activates ATF-2 by phosphorylation of Thr69 and Thr71 (2-4). Both SAPK and p38 MAPK have been shown to phosphorylate ATF-2 at these sites in vitro and in cells transfected with ATF-2. Mutations of these sites result in the loss of stress-induced transcription by ATF-2 (2-4). In addition, mutations at these sites reduce the ability of E1A and Rb to stimulate gene expression via ATF-2 (2).
ATF-7 is another member of the ATF/CREB family of leucine zipper proteins (5). Similarly, Thr51 and Thr53 (corresponding to Thr69 and Thr71 of ATF-2, respectively) can be phosphorylated under different conditions (6,7).

  1. Abdel-Hafiz, H.A. et al. (1992) Mol Endocrinol 6, 2079-89.
  2. Gupta, S. et al. (1995) Science 267, 389-93.
  3. van Dam, H. et al. (1995) EMBO J 14, 1798-811.
  4. Livingstone, C. et al. (1995) EMBO J 14, 1785-97.
  5. Peters, C.S. et al. (2001) J Biol Chem 276, 13718-26.
  6. Camuzeaux, B. et al. (2008) J Mol Biol 384, 980-91.
  7. Maekawa, T. et al. (2010) EMBO J 29, 196-208.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
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
#44167

Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb

Western Blotting Image 1: Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb Expand Image
使用 Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb(上图)、ATF-2/ATF-7 (A9G1M) Rabbit mAb #82870(中间)或 β-Actin (D6A8) Rabbit mAb #8457 (下图)对未经处理 (-) 或已经 Anisomycin #2222(25 μg/ml,30 分钟;+)处理的 HeLa 和 NIH/3T3 细胞的提取物进行蛋白质印迹法分析。
Immunoprecipitation Image 1: Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb Expand Image
对经 Anisomycin #2222(25 μg/ml,30 分钟)处理的 HeLa 细胞的提取物 Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) 蛋白进行免疫沉淀法分析。泳道 1 为 10% input,泳道 2 为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb。使用 Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb进行蛋白质印迹法分析。
Immunofluorescence Image 1: Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb Expand Image
使用 Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb(绿色)和Lamin A/C (4C11) Mouse mAb #4777(红色),对未经处理(左图)、经 Anisomycin #2222(25 μg/mL, 30 min;中间)处理或经 anisomyci 处理并用 λ 磷酸酶后处理以证实磷酸化-特异性(右图)的 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白丝用 DyLight 554 Phalloidin #13054(伪彩蓝色)标记。
Flow Cytometry Image 1: Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb Expand Image
使用 Phospho-ATF-2 (Thr69)/ATF-7 (Thr51) (E3D6B) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(蓝色,阴性)或已经 Anisomycin #2222(25 ug/ml,30 分钟;绿色,阳性)处理的 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。