The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 10 µg/ml.
Use a concentration of 2.5 - 5 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
Use at 10 µg/mg of lysate.
Use a concentration of 10 - 20 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
Nuclear 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.
Belongs to the nuclear hormone receptor family. NR3 subfamily. Contains 1 nuclear receptor DNA-binding domain.
Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
Phosphorylated 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).
Nucleus. Cytoplasm. Cell membrane. A minor fraction is associated with the inner membrane and Nucleus. Cytoplasm. Cell membrane. Associated with the inner membrane via palmitoylation.
ICC/IF image of ab75635 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab75635 at 10µg/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 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody (ab75635)This image is courtesy of an Abreview submitted by Raísa Brito.
Immunohistochemical analysis of Capybara testis tissue, labeling Estrogen receptor alpha with ab75635, diluted 1/1000. Heat mediated antigen retrieval with Citrate buffer pH 6, blocked with BSA.