Overview

  • Product name
    Anti-Estrogen Receptor alpha antibody
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Rabbit polyclonal to Estrogen Receptor alpha
  • Tested applications
    Suitable for: ICC/IF, WB, Flow Cyt, ICC, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig, Deer
    Predicted to work with: Hamster, Cow, Cat
  • Immunogen

    Synthetic peptide corresponding to Human Estrogen Receptor alpha aa 21-32.
    Sequence:

    NELEPLNRPQLK


    Database link: P03372
    (Peptide available as ab5847)

  • Positive control
    • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab3575 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF Use a concentration of 0.5 mg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 64 kDa (predicted molecular weight: 68 kDa).Can be blocked with Human Estrogen Receptor alpha peptide (ab5847).
Flow Cyt Use at an assay dependent concentration. PubMed: 23768049
ICC Use at an assay dependent concentration.
IHC-P Use a concentration of 1 - 5 µg/ml.

Target

  • Function
    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.
  • Sequence similarities
    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • Domain
    Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • Post-translational
    modifications
    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).
  • Cellular localization
    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.
  • Information by UniProt
  • Database links
  • Alternative names
    • DKFZp686N23123 antibody
    • ER alpha antibody
    • ER antibody
    • ER-alpha antibody
    • Era antibody
    • ESR antibody
    • ESR1 antibody
    • ESR1_HUMAN antibody
    • ESRA antibody
    • Estradiol receptor antibody
    • Estrogen nuclear receptor alpha antibody
    • Estrogen receptor 1 antibody
    • Estrogen receptor alpha 3*,4,5,6,7*/822 isoform antibody
    • Estrogen receptor alpha antibody
    • Estrogen receptor alpha delta 3*,4,5,6,7*,8*/941 isoform antibody
    • Estrogen receptor alpha delta 3*,4,5,6,7*/819 2 isoform antibody
    • Estrogen receptor alpha delta 4 +49 isoform antibody
    • Estrogen receptor alpha delta 4*,5,6,7*/654 isoform antibody
    • Estrogen receptor alpha delta 4*,5,6,7,8*/901 isoform antibody
    • Estrogen receptor alpha E1 E2 1 2 antibody
    • Estrogen receptor alpha E1 N2 E2 1 2 antibody
    • Estrogen receptor antibody
    • ESTRR antibody
    • NR3A1 antibody
    • Nuclear receptor subfamily 3 group A member 1 antibody
    see all

Images

  • 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.

  • 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.

  • 4% paraformaldehyde fixed 0.1% Triton permeabilized T-47D (human ductal breast epithelial tumor cell line) cells stained for Estrogen Receptor alpha (green) using ab3575 (1/2000) in ICC/IF. DyLight 488-conjugated goat anti-rabbit IgG at 1/500.

References

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 Cancer 13:289 (2013). Flow Cyt ; Human . Read more (PubMed: 23768049) »
  • Fu S  et al. Aurora kinase inhibitor VE 465 synergistically enhances cytotoxicity of carboplatin in ovarian cancer cells through induction of apoptosis and downregulation of histone 3. Cancer Biol Ther 13:1034-41 (2012). Read more (PubMed: 22895067) »

See all 5 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast Cancer Metastasis in Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: ph6 citrate
Permeabilization
No
Specification
Breast Cancer Metastasis in Lung
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Formaldehyde
Username

Dr. Victoria Thompson

Verified customer

Submitted May 13 2016

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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