Product nameAnti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade
See all Estrogen Receptor alpha primary antibodies
DescriptionMouse monoclonal [C-542] to Estrogen Receptor alpha - ChIP Grade
SpecificityThis affinity purified antibody identifies recombinant human estrogen receptor. Binding of antibody to estrogen receptor was blocked by excess free peptide containing ER sequences 582- 595, but not by an unrelated peptide.
Tested applicationsSuitable for: ChIP, IP, ICC/IF, WB, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Hamster, Human
Predicted to work with: Sheep, Horse, Cow, Chimpanzee
General notesStorage below -20°C is not recommended.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferPreservative: 0.09% Sodium Azide
Constituents: 50% Glycerol, PBS, pH 7.2
Concentration information loading...
Purification notesPurified from ascites.
ChIP Related Products
Our Abpromise guarantee covers the use of ab66102 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ChIP||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.
Use 20ug/100ul beads: It is recommend that protein A Sepharose be used as immunoabsorbent with a bridging rabbit anti-mouse IgG to ensure efficient binding of the antibody
|ICC/IF||Use at an assay dependent concentration.|
|WB||Use a concentration of 2 µg/ml. Predicted molecular weight: 68 kDa.|
|EMSA||Use at an assay dependent concentration.
20ug per reaction
|Flow Cyt||Use 0.1-1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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.
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.
- Information by UniProt
- DKFZp686N23123 antibody
- ER alpha antibody
- ER antibody
Overlay histogram showing MCF7 cells stained with ab66102 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab66102, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Cross-linking (X-chIP) analysis of human MCF7 nuclear cell lysate labeling Estrogen Receptor alpha with ab66102 at concentration 5 µg/µg chromatin. Formaldehyde was used as the cross-linking agent. Semiquantitative PCR used for detection. Cells were incubated for 16 hours at 4°C with chIP dilution buffer.
Anti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade (ab66102) at 3 µg/ml + Human MCF7 whole cell lysate at 100 µg
Polyclonal Goat anti-mouse HRP at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 68 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?
Exposure time: 10 minutes
Blocking step: 5% milk for 1 hour at 37°C.
This product has been referenced in:
- Lin J et al. MiR-203 inhibits estrogen-induced viability, migration and invasion of estrogen receptor a-positive breast cancer cells. Exp Ther Med 14:2702-2708 (2017). Read more (PubMed: 28962215) »
- Li R et al. Mice endometrium receptivity in early pregnancy is impaired by maternal hyperinsulinemia. Mol Med Rep 15:2503-2510 (2017). WB ; Mouse . Read more (PubMed: 28447735) »