Product Pathways - Nuclear Receptor Signaling
Phospho-Estrogen Receptor α Antibody Sampler Kit #9924
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Isotype |
|---|---|---|---|---|---|
| Estrogen Receptor α (D8H8) Rabbit mAb #8644 | 40 µl | W IP IF-IC ChIP | H | 66 | Rabbit |
| Phospho-Estrogen Receptor α (Ser104/106) Antibody #2517 | 40 µl | W | H (M) | 66 | Rabbit |
| Phospho-Estrogen Receptor α (Ser118) (16J4) Mouse mAb #2511 | 40 µl | W IHC-P | H | 66 | Mouse IgG2b |
| Phospho-Estrogen Receptor α (Ser167) (D1A3) Rabbit mAb #5587 | 40 µl | W | H (Mk) | 66 | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody #7074 | 100 µl | Goat | |||
| Anti-mouse IgG, HRP-linked Antibody #7076 | 100 µl | Horse |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IHC-P=Immunohistochemistry (Paraffin)
IF-IC=Immunofluorescence (Immunocytochemistry)
ChIP=Chromatin IP
Reactivity Key:
H=Human
M=Mouse
Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Phospho-Estrogen Receptor α (Ser167) (D1A3) Rabbit mAb detects endogeneous levels of ERα protein only when phosphorylated at Ser167, and also cross reacts with a nonspecific band around 77 kDa. Phospho-Estrogen Receptor α (Ser104/106) Antibody detects endogenous levels of Ser104/106 phosphorylated ERα. Phospho-Estrogen Receptor α (Ser118) (16J4) Mouse mAb will detect endogenous levels of Ser118 phosphorylated ERα. Estrogen Receptor α (D8H8) Rabbit mAb 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 α, stimulated with EGF and E2, using Phospho-Estrogen Receptor α (Ser118) (16J4) Mouse mAb #2511 (upper) and control Estrogen Receptor α Antibody #2512 (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
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.)
Western Blotting
Western analysis of extracts from MCF7 cells, untreated or treated with Estrodiol/EGF (100nM each, together for 30 min) and further treated with calf intestinal phosphatase (CIP), using Phospho-Estrogen Receptor α (Ser167) (D1A3) Rabbit mAb #5587 (upper) or Estrogen Receptor α (D62A3) Mouse mAb #2512 (lower).
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 #8644 (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Description
The Phospho-Estrogen Receptor α Antibody Sampler Kit provides an economical means to evaluate the activation status of ERα, including phosphorylation of Ser104/106, Ser167, and Ser118. The monoclonal control ERα antibody is also included to detect total Estrogen Receptor α levels. The kit contains enough primary antibody to perform four Western blot experiments.
Source / Purification
Activation state monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides or non-phosphopeptide corresponding to residues surrounding Ser167, Ser118, and the carboxy terminus of human estrogen receptor α protein. Activation state polyclonal antibody is produced by immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser104/106 of human ERα. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
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).
- 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
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Protocols
- 2511:
- IHC / Paraffin, Western Blotting
- 2517:
- Western Blotting
- 5587:
- Western Blotting
- 8644:
- ChIP Agarose, ChIP Magnetic, Immunofluorescence, Immunoprecipitation, Western Blotting
Companion Products
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
