Cell Signaling Technology

Product Pathways - Nuclear Receptor Signaling

Estrogen Receptor α (D8H8) Rabbit mAb #8644

sc-514910   sc-542   sc-7207   sc-787  

No. Size Price
8644S 100 µl ( 10 western blots ) ¥3,100.00 现货查询 购买询价
8644T 20 µl ( 2 western blots ) ¥1,200.00 现货查询 购买询价
8644 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human, Endogenous 66 Rabbit IgG
IP 1:50
IF-IC 1:1600
ChIP 1:100

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), ChIP=Chromatin IP,

Specificity / Sensitivity

Estrogen Receptor α (D8H8) Rabbit mAb recognizes endogenous levels of total ERα protein.Estrogen Receptor α (D8H8) Rabbit mAb识别内源性水平的总ERα蛋白。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy terminus of human ERα protein.该单克隆抗体通过用合成肽免疫动物制备,该合成肽是人ERα蛋白靠近羧基末端的残基。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells grown in phenol red free medium and 5% charcoal stripped FBS for 4 d then treated with β-estradiol (10 nM) for 1 h and either 5 μl of Estrogen Receptor α (D8H8) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.通过交联染色质进行染色质免疫沉淀实验,交联染色质来自于4 x 106个细胞,细胞在不含酚红,含有活性炭/葡聚糖处理的5%FBS中培养4天,随后用 β-雌二醇(10 nM) 处理1 h,和 5 μl Estrogen Receptor α (D8H8) Rabbit mAb或2 μl Normal Rabbit IgG #2729处理,使用的试剂盒为SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003。通过real-time PCR 方法,对提取的DNA进行量化,使用的引物为SimpleChIP® Human ESR1 Promoter Primers #9673, SimpleChIP® Human pS2 Promoter Primers #9702和SimpleChIP® Human α Satellite Repeat Primers #4486. 在每个样品中免疫沉淀的DNA量表示为相当于加入染色质总量,相当于一个。

Western Blotting

Western Blotting

Western blot analysis of extracts from ER-positive cell lines (MCF7, T-47D, ZR-75-1) and ER-negative cell lines (SK-BR-3 and MCF 10A) using Estrogen Receptor α (D8H8) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).Western blot 方法检测ER-阳性细胞系(MCF7, T-47D, ZR-75-1)提取物和ER-阴性细胞系(SK-BR-3和MCF 10A),使用的抗体为Estrogen Receptor α (D8H8) Rabbit mAb (上图)或 β-Actin (D6A8) Rabbit mAb #8457 (下图).



Confocal immunofluorescent analysis of MCF7 (left) or SK-BR-3 (right) cells using Estrogen Receptor α (D8H8) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).激光共聚焦免疫荧光方法检测MCF7 细胞(左图)或SK-BR-3 细胞(右图),使用的抗体为 Estrogen Receptor α (D8H8) Rabbit mAb (绿色).肌动蛋白丝用DY-554鬼笔环肽标记(红色)。


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).雌激素受体α(ERα),是类固醇受体超家族的一员,含有高度保守的DNA结合域(DBD)和配体结合域(LBD)(1)。ERα通过其雌激素非依赖性和雌激素依赖性激活域(分别为AF-1和AF-2),调节转录招募共激活因子蛋白和与一般转录机制的相互作用(2)。磷酸化提供了调节ERα活性的重要机制(3,4)。 ERα有多个磷酸化位点(5)。丝氨酸(104,106,118和167位)都位于氨基末端的转录激活功能域AF-1,这些丝氨酸的磷酸化在调节ERα活性中起着重要作用。丝氨酸(118位)可能是转录调节激酶CDK7的底物(5)。丝氨酸(167位)可以通过p90RSK和Akt磷酸化(4,6)。 Ser167的磷酸化可能会使乳腺癌患者对他莫昔芬耐药(4)。

  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.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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