ChIP, C&R
H
Endogenous
Rabbit IgG
#P03372
2099
Product Information
Product Usage Information
For optimal ChIP results, use 5 μl of antibody and 10 μg of chromatin (approximately 4 x 106 cells) per IP. This antibody has been validated using SimpleChIP® Enzymatic Chromatin IP Kits.
The CUT&RUN dilution was determined using CUT&RUN Assay Kit #86652.
| Application | Dilution |
|---|---|
| Chromatin IP | 1:100 |
| CUT&RUN | 1:100 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser167 of human ERα protein.
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).
ERα can be phosphorylated at Ser167 by various kinases such as S6K1, RSK, and Aurora A (7-9). Phosphorylation on Ser167 promotes ERα-dependent transcription and cellular proliferation, and is attributed to increased resistance to tamoxifen treatment (6, 9, 10). Various studies have shown that increased Ser167 phosphorylation correlates with poor prognosis in different cancer types (11, 12)
- Mangelsdorf, D.J. et al. (1995) Cell 83, 835-9.
- Glass, C.K. and Rosenfeld, M.G. (2000) Genes Dev 14, 121-41.
- Chen, D. et al. (1999) Mol Cell Biol 19, 1002-15.
- Campbell, R.A. et al. (2001) J Biol Chem 276, 9817-24.
- Chen, D. et al. (2000) Mol Cell 6, 127-37.
- Joel, P.B. et al. (1998) Mol Cell Biol 18, 1978-84.
- Yamnik, R.L. et al. (2009) J Biol Chem 284, 6361-9.
- Yamnik, R.L. and Holz, M.K. (2010) FEBS Lett 584, 124-8.
- Zheng, X.Q. et al. (2014) Oncogene 33, 4985-96.
- Wang, Y. et al. (2015) J Mol Endocrinol 54, 351-61.
- López-Calderero, I. et al. (2014) Hum Pathol 45, 2437-46.
- Kato, E. et al. (2014) Cancer Sci 105, 1307-12.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Applications Key
ChIP: Chromatin IP C&R: CUT&RUN
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
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