Cell Signaling Technology

Product Pathways - Nuclear Receptor Signaling

Phospho-Estrogen Receptor alpha (Ser104/106) Antibody #2517

Applications Reactivity MW (kDa) Source
W IF-IC H (M) 66 Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-Estrogen Receptor alpha (Ser104/106) Antibody detects endogenous levels of ER alpha only when phosphorylated at Ser104/106. It does not cross-react with the phosphorylated ER isoform beta.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding serine 104/106 of human ER alpha. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from COS-1 cells expressing wild-type or mutant ER alpha, stimulated with beta-estradiol (100 nM) and EGF (100 ng/ml) for 30 minutes, using Phospho-Estrogen Receptor alpha (Ser104/106) Antibody (upper) or control ER alpha Antibody #2512 (lower). (Cell lysates provided by Dr. Simak Ali, Hammersmith Hospital, London.)

Western Blotting

Western Blotting

Dephosphorylation of wild-type ER alpha abolishes Phospho-Estrogen Receptor alpha (Ser104/106) Antibody's recognition.

IF-IC

IF-IC

Immunofluorescent staining of MCF-7 cells unstimulated (left) or stimulated with beta-estradiol (100 nM) and EGF (100 ng/ml) for 30 minutes (right), using Phospho-Estrogen Receptor alpha (Ser104/106) Antibody.


Background

Estrogen receptorα (ER α), a member of the steroid receptor superfamily, contains highly conserved DNA binding (DBD) and ligand binding domains (LBD) (1). Through its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively), ER α regulates transcription by recruiting coactivator proteins and interacting with general transcriptional machinery (2). Phosphorylation provides an important mechanism to regulate ER α activity (3,4). ER α is phosphorylated on multiple sites (5). Serines 104, 106, 118 and 167 are located in the amino-terminal transcription activation function domain AF-1, and phosphorylation of these serines plays an important role in regulating ER α activity. Ser118 may be the substrate of the transcription regulatory kinase cdK7 (5). Ser167 may be phosphorylated by p90RSK and Akt (4,6). Phosphorylation of Ser167 may confer tamoxifen resistance in breast cancer patients (4).

  1. Mangelsdorf, D.J. et al. (1995) Cell 83, 835-839.
  2. Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev. 14, 121-141.
  3. Chen, D. et al. (1999) Mol. Cell. Biol. 19, 1002-1015.
  4. Campbell, R.A. et al. (2001) J. Biol. Chem. 276, 9817-9824.
  5. Chen, D. et al. (2000) Mol. Cell 6, 127-137.
  6. Joel, P.B. et al. (1998) Mol. Cell. Biol. 18, 1978-1984.

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