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Technical Data Sheet

EK6720

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Estrogen Receptor Phospho-Regulation
Antibody Sampler Kit
Price

$405

Kit Components
Applications: WB = Western blot, E = ELISA, ICC = Immunocytochemistry, IP = Immunoprecipitation, IHC = Immunohistochemistry Species: H = Human, R = Rat, M = Mouse, C = Chicken, F = Fish

Product References
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
MS3001 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot
Kit Summary
The ERα Phospho-Regulation antibody sampler kit can be used to detect phosphorylation of Tyr-537 relative to total ERα expression levels. The kit includes mouse monoclonal and rabbit polyclonal antibodies to detect phospho-Tyr-537, as well as a rabbit polyclonal antibody to detect ERα. The kit also includes anti-Rabbit Light Chain specific:HRP and anti-Mouse Ig specific:HRP secondary reagents for detection of antibodies in Western blot, ELISA, or immunocytochemistry.
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Western blot image of human MCF-7 cells treated with pervanadate (1 mM) for 30 min. (lanes 1-6). The some lanes of the blot were treated with alkaline phosphatase (lanes 2, 4 & 6). The blot was probed with mouse monoclonal anti-ERα (Tyr-537) phospho-specific (lanes 1 & 2), rabbit polyclonal anti-ERα (C-terminus) (lanes 3 & 4), and rabbit polyclonal anti-ERα (Tyr-537) phospho-specific (lanes 5 & 6).

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Immunocytochemical labeling of Estrogen Receptor α in paraformaldehyde fixed and NP-40 permeabilized MDA-MB-231 cells. The cells were labeled with rabbit polyclonal anti-Estrogen Receptor α (EP5431). The antibody was detected using goat anti-rabbit DyLight® 594.

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Background
Estrogen receptor α (ERα) is a member of the steroid receptor superfamily and its structure includes highly conserved DNA binding and ligand binding domains found in this family. ERα regulates transcription at estrogen response elements by recruiting co-activator proteins and transcription proteins via its estrogen-independent and estrogen-dependent activation domains (AF-1 and AF-2, respectively). Phosphorylation at multiple sites provides an important mechanism to regulate ERα activity. Ser-104, Ser-106, Ser-118, and Ser-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. In addition to these sites, phosphorylation of Tyr-537 has been implicated in maximal hormone binding, dimerization, and transcriptional activity. Tyr-537 is phosphorylated by c-Src leading to nuclear export of ERα and degradation. Thus, a variety of phosphorylation events control ERα activity.

Background References
Castoria, G. et al. (2012) Oncogene. 31:4868.
Anbalagan M, Rowan BG (2015) Mol Cell Endocrin. 418(3):264.
Buffer and Storage
Primary antibodies are supplied in phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in the same buffer without azide. Store all at –20°C. Stable for 1 year.
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