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, IF-IC

REACTIVITY:

H

SENSITIVITY:

Endogenous

MW (kDa):

66

Source/Isotype:

Rabbit IgG

UniProt ID:

#P03372

Entrez-Gene Id:

2099

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 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-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb recognizes endogenous levels of ERα protein only when phosphorylated at Ser167.

Species Reactivity:

Human

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser167 of human ERα protein.

Background

Estrogen receptor α (ERα), a member of the steroid receptor superfamily, contains highly conserved DNA binding and ligand binding domains (1). Through its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively), ERα regulates transcription by recruiting coactivator proteins and interacting with general transcriptional machinery (2). Phosphorylation at multiple sites provides an important mechanism to regulate ERα activity (3-5). Ser104, 106, 118, and 167 are located in the amino-terminal transcription activation function domain AF-1, and phosphorylation of these serine residues plays an important role in regulating ERα activity. Ser118 may be the substrate of the transcription regulatory kinase CDK7 (5). Ser167 may be phosphorylated by p90RSK and Akt (4,6). According to the research literature, phosphorylation at Ser167 may confer tamoxifen resistance in breast cancer patients (4).
ERα can be phosphorylated at Ser167 by various kinases such as S6K1, RSK, and Aurora A (7-9). Phosphorylation on Ser167 promotes ERα-dependent transcription and cellular proliferation, and is attributed to increased resistance to tamoxifen treatment (6, 9, 10). Various studies have shown that increased Ser167 phosphorylation correlates with poor prognosis in different cancer types (11, 12)

  1. Mangelsdorf, D.J. et al. (1995) Cell 83, 835-9.
  2. Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev 14, 121-41.
  3. Chen, D. et al. (1999) Mol Cell Biol 19, 1002-15.
  4. Campbell, R.A. et al. (2001) J Biol Chem 276, 9817-24.
  5. Chen, D. et al. (2000) Mol Cell 6, 127-37.
  6. Joel, P.B. et al. (1998) Mol Cell Biol 18, 1978-84.
  7. Yamnik, R.L. et al. (2009) J Biol Chem 284, 6361-9.
  8. Yamnik, R.L. and Holz, M.K. (2010) FEBS Lett 584, 124-8.
  9. Zheng, X.Q. et al. (2014) Oncogene 33, 4985-96.
  10. Wang, Y. et al. (2015) J Mol Endocrinol 54, 351-61.
  11. López-Calderero, I. et al. (2014) Hum Pathol 45, 2437-46.
  12. Kato, E. et al. (2014) Cancer Sci 105, 1307-12.

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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry)

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.
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。

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Revision 1
#64508

Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb

Western Blotting Image 1: Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb Expand Image
对未经 (-) 或已经以下联合处理方法按规定处理的MCF7 细胞提取物进行蛋白质印迹分析:用 Human Epidermal Growth Factor (hEGF) #8916(100ng/ml,15 分钟)或 hEGF 处理,随后再用牛小肠磷酸酶 (CIP) 和 λ 磷酸酶处理,使用 Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb(上图)或 Estrogen Receptor α (D8H8) Rabbit mAb #8644(下图)。
Immunofluorescence Image 1: Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb Expand Image
使用 Phospho-Estrogen Receptor α (Ser167) (D5W3Z) Rabbit mAb(绿色),对未处理的(左上图)、经 Human Epidermal Growth Factor (hEGF) #8916(10 ng/mL,30 分钟;右上图)刺激或经 hEGF 刺激后再使用 λ 磷酸酶后处理(左下图)的血清饥饿 MCF7 细胞(ERα 阳性)进行共聚焦免疫荧光分析。经 hEGF 处理的 SK-BR-3 细胞(ERα 阴性;右下图)包括在内,作为附加阴性对照。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。