ICC: Human coronary artery smooth muscle cells (See Han et al). IHC-P: Rat prostate tissue. ICC/IF: T-47D cells.
This antibody gave a positive result in IHC in the following FFPE tissue: Human breast ductal carcinoma
This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HepG2.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Use at an assay dependent concentration. PubMed: 23768049
Use at an assay dependent concentration.
Use a concentration of 1 - 5 µg/ml.
FunctionNuclear hormone receptor. The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Can activate the transcriptional activity of TFF1.
Sequence similaritiesBelongs to the nuclear hormone receptor family. NR3 subfamily. Contains 1 nuclear receptor DNA-binding domain.
DomainComposed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
Post-translational modificationsPhosphorylated by cyclin A/CDK2. Phosphorylation probably enhances transcriptional activity. Glycosylated; contains N-acetylglucosamine, probably O-linked. Ubiquitinated. Deubiquitinated by OTUB1. Dimethylated by PRMT1 at Arg-260. The methylation may favor cytoplasmic localization. Palmitoylated (isoform 3). Not biotinylated (isoform 3).
Cellular localizationNucleus. Cytoplasm. Cell membrane. A minor fraction is associated with the inner membrane and Nucleus. Cytoplasm. Cell membrane. Associated with the inner membrane via palmitoylation.
ab3575 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 minutes) and then incubated in 1% BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab3575 at 5 µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 dilution for 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.
IHC image of Estrogen Receptor alpha staining in human breast ductal carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab3575, 5 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ab3575 at a dilution of 5 µg/ml staining ER alpha in rat prostate tissue sections.
Western blot - Anti-Estrogen Receptor alpha antibody (ab3575)
All lanes : Anti-Estrogen Receptor alpha antibody (ab3575) at 1/1000 dilution
Lane 1 : Recombinant full-length human Estrogen Receptor alpha protein, 250 ng Lane 2 : Recombinant full-length human Estrogen Receptor alpha protein, 125 ng Lane 3 : Recombinant full-length human Estrogen Receptor alpha protein, 62.5 ng Lane 4 : Recombinant full-length human Estrogen Receptor alpha protein, 31.25 ng
Secondary HRP-conjugated goat anti-rabbit IgG at 1/40000 dilution Developed using the ECL technique
Predicted band size : 68 kDa
Blocked with 5% milk in TBST for at least 1 hour at room temperature.
References for Anti-Estrogen Receptor alpha antibody (ab3575)
This product has been referenced in:
Ghebeh H et al. Profiling of normal and malignant breast tissue show CD44high/CD24low phenotype as a predominant stem/progenitor marker when used in combination with Ep-CAM/CD49f markers. BMC Cancer13:289 (2013).
Read more (PubMed: 23768049) »
Anagnostis A & Papadopoulos AI Effects of a diet rich in sesame ( Sesamum indicum) pericarp on the expression of oestrogen receptor alpha and oestrogen receptor beta in rat prostate and uterus. Br J Nutr102:703-8 (2009).
Read more (PubMed: 19309532) »