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

Overview

  • Product nameAnti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Rabbit monoclonal [EPR4097] to Estrogen Receptor alpha - ChIP Grade
  • SpecificityFor Mouse, ab108398 is recommended for WB only. Rat cross reactivity tested by WB only.
  • Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ChIP, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Estrogen Receptor alpha aa 1-300. Recombinant Human Estrogen Receptor alpha protein.

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

    This product is a recombinant rabbit monoclonal antibody.

    A trial size is available to purchase for this antibody.

    Alternative versions available:

    Anti-Estrogen Receptor alpha antibody (Alexa Fluor® 488) [EPR4097] (ab205850)

    Anti-Estrogen Receptor alpha antibody (Alexa Fluor® 647) [EPR4097] (ab205851)

    Anti-Estrogen Receptor alpha antibody (Alexa Fluor® 594) [EPR4097] (ab207046)

    Anti-Estrogen Receptor alpha antibody (Alexa Fluor® 568) [EPR4097] (ab207261)

     

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    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.

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 protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

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

  • 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.
  • 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 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
  • 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

Anti-Estrogen Receptor alpha antibody [EPR4097] - ChIP Grade images

  • 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
    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.

  • 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.

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

ab108398 has not yet been referenced specifically in any publications.

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Pig Cell lysate - whole cell (Pig Mammary Epithelial Cells)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 15 µg
Specification Pig Mammary Epithelial Cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 10 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Spodoptera frugiperda (SF9 cells) Cell lysate - whole cell (sf9 cells)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 30 nM Bortezomib for 16 hours
Specification sf9 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 10 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Rat Cell lysate - whole cell (rat intestinal epithelial cells (RIE-1))
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 100 nM Bortezomib for 16 hours
Specification rat intestinal epithelial cells (RIE-1)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 10 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Chinese Hamster Cell lysate - whole cell (Chinese hamster ovary cell)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 100 nM Bortezomib for 16 hours
Specification Chinese hamster ovary cell
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Feb 10 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (rat intestinal epithelial cells (RIE-1))
Permeabilization Yes - 0.2% TritonX-100
Specification rat intestinal epithelial cells (RIE-1)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Feb 09 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Chinese Hamster Cell (Chinese hamster ovary cell)
Permeabilization Yes - 0.2% TritonX-100
Specification Chinese hamster ovary cell
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
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Submitted Feb 09 2016

Application Flow Cytometry
Sample Mouse Cell (Mouse Lewis Lung Carcinoma cells)
Permeabilization Yes - 0.2% Triton X-100
Gating Strategy No gating
Specification Mouse Lewis Lung Carcinoma cells
Preparation Cell harvesting/tissue preparation method: trypsin
Sample buffer: Hanks’ Balanced Salt solution (HBSS)
Fixation Paraformaldehyde
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Submitted Jan 21 2016

Application IHC - Wholemount
Sample Human Tissue (Invasive ductal carcinoma)
Specification Invasive ductal carcinoma
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Submitted Dec 19 2015

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (COS-7 cells)
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Loading amount 100000 cells
Treatment 30 nM Bortezomib for 24 hours
Specification COS-7 cells
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted Jun 26 2015

Application Western blot
Sample Mouse Cell lysate - whole cell (Lewis lung cancer, FL5.12, mouse embryonic fibrobl)
Gel Running Conditions Reduced Denaturing (4-20% Bis-Tris)
Loading amount 100000 cells
Specification Lewis lung cancer, FL5.12, mouse embryonic fibrobl
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
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Submitted May 11 2015

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