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 # PA1-308) at a dilution of 1:4000 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, Rat|
|Published species reactivity||Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues M(1) T M T L H T K A S G M A L L H Q I Q G N(21) 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 (Frozen) (IHC (F))||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.
PA1-308 has been successfully used in Western blot, immunofluorescence and immunohistochemistry procedures. By Western blot, this antibody detects an ~64 kDa protein representing endogenous levels of ER alpha from MCF-7 cell extracts. Immunohistochemical staining of ER alpha in rat prostate with PA1-308 results in primarily nuclear staining.
The PA1-308 immunogen is a synthetic peptide corresponding to residues M(1) T M T L H T K A S G M A L L H Q I Q G N(21) C of human ER alpha. This peptide sequence is completely conserved between human, rat and mouse and is highly conserved between sheep, porcine, camel, bovine and chicken ER alpha. PA1-308 immunizing peptide (Cat. # PEP-036) 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.
Characterization of bladder sensory neurons in the context of myelination, receptors for pain modulators, and acute responses to bladder inflammation.
PA1-308 was used in immunohistochemistry to study bladder neurons involved in pain modulation and acute responses to inflammation
|Forrest SL,Osborne PB,Keast JR||Frontiers in neuroscience (7:null)||2013|
Pretreatment with phytoestrogen-rich plant decreases breast tumor incidence and exhibits lower profile of mammary ERalpha and ERbeta.
PA1-308 was used in immunohistochemistry to investigate the protective effect of Pueraria mirifica phytoestrogens against rat mammary carcinoma tumors induced by DMBA
|Cherdshewasart W,Panriansaen R,Picha P||Maturitas (58:174)||2007|
Androgen and estrogen receptor-mediated mechanisms of testosterone action in male rat pelvic autonomic ganglia.
PA1-308 was used in immunohistochemistry to investigate the mechanisms of testosterone action in male rat pelvic autonomic ganglia
|Purves-Tyson TD,Arshi MS,Handelsman DJ,Cheng Y,Keast JR||Neuroscience (148:92)||2007|
Sex difference in septal neurons projecting axons to midbrain central gray in rats: a combined double retrograde tracing and ER-immunohistochemical study.
PA1-308 was used in immunohistochemistry to investigate the sexual dimorphism in septal neurons projecting axons to midbrain central gray in rats.
|Tsukahara S,Yamanouchi K||Endocrinology (143:285)||2002|
Cancer-associated myofibroblasts possess various factors to promote endometrial tumor progression.
PA1-308 was used in immunohistochemistry to demonstrate the role of cancer-associated myofibroblasts in endometrial tumor progression.
|Orimo A,Tomioka Y,Shimizu Y,Sato M,Oigawa S,Kamata K,Nogi Y,Inoue S,Takahashi M,Hata T,Muramatsu M||Clinical cancer research : an official journal of the American Association for Cancer Research (7:3097)||2001|
Differential effects of steroids on the synthesis of polyunsaturated fatty acids by human neuroblastoma cells.
PA1-308 was used in western blot to investigate the effects of steroids on polyunsaturated fatty acid synthesis by human neuroblastoma cells
|Extier A,Perruchot MH,Baudry C,Guesnet P,Lavialle M,Alessandri JM||Neurochemistry international (55:295)||2009|
Genistein affects HER2 protein concentration, activation, and promoter regulation in BT-474 human breast cancer cells.
PA1-308 was used in western blot to study the effect of genistein on HER2 expression and function in BT-474 human breast cancer cells
|Sakla MS,Shenouda NS,Ansell PJ,Macdonald RS,Lubahn DB||Endocrine (32:69)||2007|
Phytosterol Pygeum africanum regulates prostate cancer in vitro and in vivo.
PA1-308 was used in western blot to investigate the effect of phytosterol Pygeum africanum on prostate cancer
|Shenouda NS,Sakla MS,Newton LG,Besch-Williford C,Greenberg NM,MacDonald RS,Lubahn DB||Endocrine (31:72)||2007|
Apoptotic and anti-proliferative effects of 17beta-estradiol and 17beta-estradiol-like compounds in the Hep3B cell line.
PA1-308 was used in western blot to study the apoptotic and anti-proliferative effects of estrogen and estrogen-like compounds on the human hepatoma Hep3B cell line
|Huang EJ,Wu CC,Huang HP,Liu JY,Lin CS,Chang YZ,Lin JA,Lin JG,Chen LM,Lee SD,Kuo WW,Huang CY||Molecular and cellular biochemistry (290:1)||2006|
Phytoestrogens in common herbs regulate prostate cancer cell growth in vitro.
PA1-308 was used in western blot to investigate the effect of phytoestrogens on prostate tumor growth
|Shenouda NS,Zhou C,Browning JD,Ansell PJ,Sakla MS,Lubahn DB,Macdonald RS||Nutrition and cancer (49:200)||2004|
Multiple forms of estrogen receptor-alpha in cerebral blood vessels: regulation by estrogen.
PA1-308 was used in western blot to investigate the expression, localization, and regulation of ER-alpha in female rat cerebral vasculature
|Stirone C,Duckles SP,Krause DN||American journal of physiology. Endocrinology and metabolism (284:E184)||2003|
17-Beta-estradiol regulates expression of genes that function in macrophage activation and cholesterol homeostasis.
PA1-308 was used in western blot to investigate the regulation of 17-beta-estradiol on the expression of genes functioning in macrophage activation and cholesterol metabolism
|Kramer PR,Wray S||The Journal of steroid biochemistry and molecular biology (81:203)||2002|
Rapid actions of estradiol on cyclic amp response-element binding protein phosphorylation in dorsal root ganglion neurons.
PA1-308 was used in immunocytochemistry to investigate the effect of estrogens on the CREB-mediated transcriptional regulation in sensory neurons
|Purves-Tyson TD,Keast JR||Neuroscience (129:629)||2004|
Liganded androgen receptor interaction with beta-catenin: nuclear co-localization and modulation of transcriptional activity in neuronal cells.
PA1-308 was used in immunocytochemistry to investigate the association between beta-catenin and androgen receptor.
|Pawlowski JE,Ertel JR,Allen MP,Xu M,Butler C,Wilson EM,Wierman ME||The Journal of biological chemistry (277:20702)||2002|