The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa).
1/50 - 1/100.
Is unsuitable for Flow Cyt or IP.
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.
Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling Estrogen Receptor alpha (phospho S118) with ab32396 at 5 μg/ml (1/200 dilution). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, an AlexaFluor®488 Goat anti-Rabbit was used as the secondary antibody at 2 μg/ml (1/1000 dilution). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)was used to counterstain at 2.5 μg/ml (1/200 dilution). DAPI nuclear counterstain. Confocal image showing the signal increased after EGF (100ng/ml, 5 min) treatment and decreased after Lambda Protein Phosphatase treatment (31°C for 2 hours).
Western blot - Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396)
All lanes : Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396) at 1/1000 dilution
Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor. Whole cell lysates Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor. Whole cell lysates. Then the membrane was incubated with phosphatase.
Lysates/proteins at 15 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Dot blot analysis of Lane 1: Estrogen Receptor alpha (pS118) phospho peptide and Lane 2: Estrogen Receptor alpha non-phospho peptide labeling Estrogen Receptor alpha (phospho S118) with ab32396 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.
Ferraiuolo RM et al. The cyclin-like protein, SPY1, regulates the ERa and ERK1/2 pathways promoting tamoxifen resistance. Oncotarget8:23337-23352 (2017).
Read more (PubMed: 28423577) »
Yan X et al. Ginsenoside Rd. promotes non-amyloidogenic pathway of amyloid precursor protein processing by regulating phosphorylation of estrogen receptor alpha. Life SciN/A:N/A (2016).
Read more (PubMed: 27825720) »