Immunofluorescent analysis of Estrogen Receptor alpha (ER alpha, green) in T47D and Hs 578T breast cancer cells. The cells were fixed with 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 15 minutes at room temperature, and blocked with 0.3% BSA in PBS for at least 30 minutes at room temperature. Cells were stained with an ER alpha polyclonal antibody (Product # PA1-309) at a dilution of 1:2000 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 temperature. Nuclei (blue) were stained with DAPI (Product # 62247). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.
|Tested species reactivity||Human, Pig, Rat|
|Published species reactivity||Pig, Deer, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues N(21) E L E P L N R P Q L K(32) C of human ER alpha.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1-5 µg/ml|
|Western Blot (WB)||1 µg/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.
PA1-309 detects estrogen receptor (ER) alpha from human, porcine and rat tissues and cells.
PA1-309 has been successfully used in Western blot, immunofluoresence and immunohistochemistry procedures. By Western blot, this antibody detects an ~64 kDa protein representing ER alpha from MCF-7 cell extracts. Immunohistochemical staining of ER alpha in rat prostate with PA1-309 results in primarily nuclear staining.
The PA1-309 immunogen is a synthetic peptide corresponding to residues N(21) E L E P L N R P Q L K(32) C of human ER alpha. This peptide sequence is completely conserved between human, rat, mouse, rabbit, sheep, porcine and bovine species. PA1-309 immunizing peptide (Cat. # PEP-037) is available for use in neutralization and control experiments.
Recombinant ER alpha expressed in baculovirus system (Cat. # RP-310) can be purchased for control experiments in Western blot and gel shift assays.
The Estrogen Receptor alpha (ER alpha) protein is approximately 66kD, though multiple isoforms ranging from 36-66kD have been described. The ER alpha gene (ESR1) consists of more than 140kb of genomic DNA divided into 8 exons, being translated into a protein with six functionally discrete domains, labeled A through F. Each domain is functionally distinct, required for transcriptional activation, binding to estrogen response elements (ERE) in DNA, constitutive dimerization, binding to heat shock proteins, and ligand recognition. 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Immunohistochemical localization of the estrogen receptor alpha (ERalpha) and progesterone receptor (PR) in the uterus of sika deer (Cervus nippon) during pregnancy.
PA1-309 was used in immunohistochemistry to investigate the distribution of estrogen receptor alpha progesterone receptor in the uterus of pregnant deer
|Yanagawa Y,Matsuura Y,Suzuki M,Katagiri S,Tsubota T||The Japanese journal of veterinary research (56:139)||2008|
Estrogen receptors in the uterus and ovarian follicles of gilts treated with dihydrotestosterone.
PA1-309 was used in immunohistochemistry to investigate the expression of estrogen receptor alpha and beta in the uterus and ovarian follicles
|Cárdenas H,Pope WF||Domestic animal endocrinology (29:523)||2005|
Estrogen receptors alpha and beta in choroid plexus epithelial cells in Alzheimer's disease.
PA1-309 was used in immunohistochemistry to investigate the effect of estrogen on Alzheimer disease and its mechanism
|Hong-Goka BC,Chang FL||Neuroscience letters (360:113)||2004|
Exposure to the environmental estrogen bisphenol A differentially modulated estrogen receptor-alpha and -beta immunoreactivity and mRNA in male mouse testis.
PA1-309 was used in immunohistochemistry to investigate the influence of bisphenol A on estrogen receptor alpha/beta expression in male mouse testis
|Takao T,Nanamiya W,Nazarloo HP,Matsumoto R,Asaba K,Hashimoto K||Life sciences (72:1159)||2003|
ICAM-1 expression in vaginal cells as a potential biomarker for inflammatory response.
PA1-309 was used in western blot to investigate ICAM-1 as a biomarker for inflammatory response in vaginal cells
|Youn H,Hong K,Yoo JW,Lee CH||Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals (13:257)||2008|
Cell cycle regulation of breast cancer cells through estrogen-induced activities of ERK and Akt protein kinases.
PA1-309 was used in western blot to study the roles of ERK and Akt in estrogen-induced breast cancer cell proliferation
|Geffroy N,Guédin A,Dacquet C,Lefebvre P||Molecular and cellular endocrinology (237:11)||2005|
Nongenomic, endothelium-independent effects of estrogen on human coronary smooth muscle are mediated by type I (neuronal) NOS and PI3-kinase-Akt signaling.
PA1-309 was used in immunocytochemistry to study how nongenomic estrogen signal is transduced in human coronary artery smooth muscle cells
|Han G,Ma H,Chintala R,Miyake K,Fulton DJ,Barman SA,White RE||American journal of physiology. Heart and circulatory physiology (293:H314)||2007|