Product Pathways - Nuclear Receptor Signaling
Phospho-Estrogen Receptor α Antibody Sampler Kit #9924
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|---|---|
| Estrogen Receptor α (62A3) Mouse mAb # 2512 | 40 microliters | W | H | 66 | Mouse |
| Phospho-Estrogen Receptor alpha (Ser118) Antibody # 2515 | 40 microliters | W | H | 66 | Rabbit |
| Phospho-Estrogen Receptor alpha (Ser104/106) Antibody # 2517 | 40 microliters | W IF-IC | H (M) | 66 | Rabbit |
| Phospho-Estrogen Receptor alpha (Ser118) (16J4) Mouse mAb # 2511 | 40 microliters | W IHC-P | H | 66 | Mouse |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 100 microliters | Goat | |||
| Anti-mouse IgG, HRP-linked Antibody # 7076 | 50 microliters | Horse |
Applications Key:
W=Western Blotting
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:
H=Human
M=Mouse
Specificity / Sensitivity
Phospho-Estrogen Receptor α (Ser104/106) Antibody #2517 detects endogenous levels of Ser104/106 phosphorylated ERα. Both Phospho-Estrogen Receptor α (Ser118) (16J4) Monoclonal Antibody #2511 and Phospho-Estrogen Receptor α (Ser118) Antibody #2515 detect endogenous levels of Ser118 phosphorylated ERα. Estrogen Receptor α (62A3) Monoclonal Antibody #2512 detects endogenous levels of ERα. Each antibody in the kit does not cross-react with phosphorylated or nonphosphorylated ER isoforms β or other related family members.
Western Blotting
Western blot analysis of extracts from MCF-7 cells, and COS-1 cells transfected with Wild-type and mutant ER alpha, stimulated with EGF and E2, using Phospho-Estrogen Receptor alpha (Ser118) (16J4) Monoclonal Antibody #2511 (upper) and control estrogen receptor alpha antibody (lower).
Western Blotting
Western blot analysis of extracts from 293 cells expressed Wild-type ER alpha and MCF-7 cells, using Estrogen Receptor alpha 62A3 Monoclonal Antibody #2512.
Western Blotting
Western blot analysis of MCF-7 cell extracts treated with or without EGF (100 ng/ml) and E2 (10-7M) for 30 minutes, using Phospho-Estrogen Receptor alpha (Ser118) Antibody #2515 (upper) and control ER alpha antibody (lower).
Western Blotting
Western blot analysis of COS-1 cell extracts expressed Wild-type ER alpha and mutants (S106A, S118A and S167A) treated with EGF (100 ng/ml) and E2 (10-7M) for 30 minutes, using Phospho-Estrogen Receptor alpha (Ser118) Antibody #2515 (upper) and control ER alpha antibody (lower).
Western Blotting
Dephosphorylation of Wild-type ER alpha abolished Phospho-Estrogen Receptor alpha (Ser104/106) Antibody's (#2517) recognition.
Western Blotting
Western blot analysis of extracts from COS-1 cells expressed Wild-type ER alpha and mutants and stimulated with beta-estradiol (100 nM) and EGF (100 ng/ml) for 30 minutes, using Phospho-Estrogen Receptor alpha (Ser104/106) Antibody #2517 (upper) and control ER alpha antibody (lower). (Cell lysates provided by Dr. Simak Ali, Hammersmith Hospital, London.)
IHC-P (paraffin)
Immunohistochemical staining of phosphorylated estrogen receptor alpha in paraffin-embedded human breast carcinoma showing nuclear localization, using Phospho-Estrogen Receptor alpha (Ser118) 16J4 Monoclonal Antibody #2511.
IHC-F (frozen)
Immunocytochemical staining of MCF-7 cells using Estrogen Receptor alpha 62A3 Monoclonal Antibody #2512 (left) or this antibody pretreated with ER alpha-specific peptides (right).
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 #2517.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with synthetic phospho-peptides (KLH coupled) corresponding to residues surrounding Ser104/106 and 118 of human ERα. The monoclonal antibodies (#2511 and #2512) are produced by immunizing mice with synthetic nonphospho- and phospho-peptides (KLH coupled) corresponding to residues surrounding Ser118 of human ERα, respectively. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are supplied in HEPES buffer with 50% glycerol and less than 0.02% sodium azide.
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).
- 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.
Application References
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