Product Pathways - Nuclear Receptor Signaling
Estrogen Receptor α (D8H8) Rabbit mAb #8644
|W IP IF-IC ChIP||H||Endogenous||66||Rabbit IgG|
Reactivity Key: H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Estrogen Receptor α (D8H8) Rabbit mAb recognizes endogenous levels of total ERα protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy terminus of human ERα protein.
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).
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).
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 45 minutes 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.
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).
- Mangelsdorf, D.J. et al. (1995) Cell 83, 835-839.
- Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev. 14, 121-141.
- Chen, D. et al. (1999) Mol. Cell. Biol. 19, 1002-1015.
- Campbell, R.A. et al. (2001) J. Biol. Chem. 276, 9817-9824.
- Chen, D. et al. (2000) Mol. Cell 6, 127-137.
- Joel, P.B. et al. (1998) Mol. Cell. Biol. 18, 1978-1984.
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- 2511 Phospho-Estrogen Receptor α (Ser118) (16J4) Mouse mAb
- 2517 Phospho-Estrogen Receptor α (Ser104/106) Antibody
- 9002 SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
- 9003 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9004 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads)
- 9005 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9006 ChIP-Grade Protein G Magnetic Beads
- 9007 ChIP-Grade Protein G Agarose Beads
- 9673 SimpleChIP® Human ESR1 Promoter Primers
- 9702 SimpleChIP® Human pS2 Promoter Primers
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 7727 Biotinylated Protein Ladder Detection Pack
For Research Use Only. Not For Use In Diagnostic Procedures.