Recombinant
RabMAb

Anti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade (ab108398)

Overview

  • Product name
    Anti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Rabbit monoclonal [EPR4097] to Estrogen Receptor alpha - ChIP Grade
  • Host species
    Rabbit
  • Specificity
    Expression levels of ER alpha protein vary with sample type.
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ChIP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human Estrogen Receptor alpha aa 1-300. The exact sequence is proprietary.

  • Positive control
    • WB: MCF7 and T47-D cell lysates. IHC-P: Human breast ductal infiltrating carcinoma and normal breast tissues. ICC/IF: MCF-7 cells.
  • General notes

    A trial size is available to purchase for this antibody.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108398 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
WB 1/1000 - 1/10000. Predicted molecular weight: 66 kDa.

For unpurified use at 1/200 - 1/1000.

IHC-P 1/250.

See IHC antigen retrieval protocols.

For unpurified use at 1/30.

Flow Cyt Use at an assay dependent concentration.
ChIP Use at an assay dependent concentration.
ICC/IF 1/250.

For unpurified use at 1/30.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue labelling Estrogen Receptor alpha with purified ab108398 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

  • All lanes : Anti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade (ab108398) at 1/2000 dilution

    Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell). Whole cell lysates
    Lane 2 : T-47D (human mammary gland ductal carcinoma epithelial cell). Whole cell lysates
    Lane 3 : MDA-MB231 (Human breast adenocarcinoma epithelial cell) Whole cell lysates (Negative control)
    Lane 4 : HepG2 (Human hepatocellular carcinoma epithelial cell) Whole cell lysates (Negative control)
    Lane 5 : Human uterus whole tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 66 kDa
    Observed band size: 68 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    Blocking and diluting buffer: 5% NFDM/TBST

  • Anti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade (ab108398) at 1/1000 dilution + GST-Tagged Recombinant Human Estrogen Receptor beta (aa 1 to 323) protein 100ng

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 66 kDa


    Exposure time: 180 seconds


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    Lane 1: Recombinant Human Estrogen Receptor beta (aa 1 to 323) protein, (Cat#: ab158385)
    There is no cross-reactivity between ab108398 and ER beta protein.

  • All lanes : purified at 1/2000 dilution

    Lane 1 : MCF-7 cell lysate
    Lane 2 : T47-D cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

    Predicted band size: 66 kDa
    Observed band size: 67 kDa why is the actual band size different from the predicted?



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunocytochemsitry/Immunofluorescence analysis of MCF-7 cells labelling Estrogen Receptor alpha (green) with purified ab108398 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

  • ab108398 staining Estrogen Receptor alpha in the human cell line MCF-7(human breast carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • ChIP analysis using unpurified ab108398 binding Estrogen Receptor alpha in MCF7 cells derived from Human breast carcinoma. Cells were cross-linked for 10 minutes with 1% formaldehyde. Samples were incubated with undiluted primary antibody for 16 hours at 4°C. Protein binding was detected using real-time PCR.
    Positive control: Estrogen treated MCF7 cells tested at PS2 promoter.
    Negative Control:IgG ChIP and ethanol-depleted cells tested at PS2 promoter.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast ductal infiltrating carcinoma tissue labelling Estrogen Receptor alpha with unpurified ab108398.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human normal tonsil tissue. Unpurified ab108398 shows negative staining.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human colonic adenocarcinoma tissue. Unpurified ab108398 shows negative staining.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human lung adenocarcinoma tissue. Unpurified ab108398 shows negative staining.

  • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human cervical carcinoma tissue. Unpurified ab108398 shows negative staining.

References

This product has been referenced in:
  • Ameen GI & Mora S Cbl downregulation increases RBP4 expression in adipocytes of female mice. J Endocrinol 236:29-41 (2018). Read more (PubMed: 29114012) »
  • Papachristou EK  et al. A quantitative mass spectrometry-based approach to monitor the dynamics of endogenous chromatin-associated protein complexes. Nat Commun 9:2311 (2018). Read more (PubMed: 29899353) »
See all 8 Publications for this product

Customer reviews and Q&As

Answer

Thank you for your inquiry. This antibody is tested and guaranteed for ICC. We did not test this antibody for IF but it should be suitable if the correct secondary antibody (anti rabbit that is tested for IF) is used. I hope this information was helpful. Please do not hesitate to contact me again with any further questions.

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

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