Immunofluorescent analysis of Estrogen Receptor (ER, green) in T47D and Hs 578T breast cancer cells. The cells were fixed with 4% paraformaldehyde for 15 min at room temp, permeabilized with 0.1% Triton X-100 for 15 min at room temp, and blocked with 0.3% BSA in PBS for at least 30 min at room temp. Cells were stained with an ER polyclonal antibody (Product # PA5-16440) at a dilution of 1:100 in blocking buffer overnight at 4C, and then incubated with a DyLight 488-conjugated goat anti-rabbit IgG secondary antibody (Product # 35552) at a dilution of 1:500 for at least 1 hour at room temp. Nuclei (blue) were stained with DAPI (Product # 62247). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Hamster, Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from the C-terminus of human estrogen receptor, alpha protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:62.5-1:125|
|Immunoprecipitation (IP)||10 µg/ml|
|Western Blot (WB)||2-5µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunocytochemistry (ICC)||See 1 publications below|
PA5-16440 targets Estrogen Receptor in IF, IHC (P), IP, and WB applications and shows reactivity with mouse, Hamster, and Human samples.
The PA5-16440 immunogen is a synthetic peptide derived from the C-terminus of human estrogen receptor, alpha protein.
Estrogen Receptors (ER) are members of the steroid family of nuclear receptors. There are two different forms of the estrogen receptor, alpha and beta, encoded by separate genes (ESR1 and ESR2, respectively). Each protein contains distinct functional domains required for transcriptional activation, binding to estrogen response elements (ERE) in DNA, constitutive dimerization, binding to heat shock proteins, and ligand recognition. ER is a ligand-activated transcription factor, that when bound to estrogen hormone, induces a conformational change that allows dimerization and binding to EREs. When bound to DNA, ER can positively or negatively regulate gene transcription through the recruitment of coactivator or corepressor proteins. There are several different isoforms of both ER alpha and ER beta. ER is an important regulator of growth and differentiation in the mammary gland. The presence of ER in breast tumors indicates an increased likelihood of response to anti-estrogen (e.g. tamoxifen) therapy.
IP-MS enrichment of ESR1 (LFQ intensity): ESR1 was enriched 162-fold from MCF7 lysate compared to background proteins, using the optimized IP-MS workflow with Pierce MS-Compatible Magnetic IP Kit protein A/G (Part No. 90409) and ESR1 antibody (Part No. PA5-16440). The STRING database (www.string-db.org) was used to identify the protein interactor list. See more information on IP-MS verification of antibody selectivity. IP-MS validation info.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
ER-¿36-mediated rapid estrogen signaling positively regulates ER-positive breast cancer stem/progenitor cells.
PA5-16440 was used in immunocytochemistry and western blot to study the modulation of ER-positive breast cancer stem/progenitior cell growth by ER-alpha36 rapid estrogen signaling
|Deng H,Zhang XT,Wang ML,Zheng HY,Liu LJ,Wang ZY||PloS one (9:null)||2014|