Anti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade (ab66102)

Overview

  • Product nameAnti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Mouse 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 reactivity
    Reacts with: Mouse, Rat, Hamster, Human
    Predicted to work with: Sheep, Horse, Cow, Chimpanzee
  • Immunogen

    Synthetic peptide:

    C-YYITGEAEGFPATV

    conjugated to KLH, corresponding to C terminal amino acids 582-595 of Human Estrogen Receptor alpha

  • General notesStorage below -20°C is not recommended.

Properties

Applications

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.

Application Abreviews Notes
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.

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 [C-542] - ChIP Grade images

  • 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.

References for Anti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade (ab66102)

ab66102 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 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (placenta)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification placenta
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Oct 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Dog Tissue sections (placenta)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification placenta
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Oct 21 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Cow Tissue sections (oviduct)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification oviduct
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Jul 15 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (placenta)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification placenta
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Jul 13 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Horse Tissue sections (nerve)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification nerve
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
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Submitted Jun 05 2015

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Antigen retrieval step None
Sample Mouse Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.3% Triton X-100
Fixative Paraformaldehyde
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Mr. Michael Milne

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Submitted Jun 03 2014

Application ChIP
Detection step Semiquantitative PCR
Sample Rat Cell lysate - nuclear (breast cells)
Specification breast cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde 10%
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Submitted Dec 12 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (8)
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Dec 09 2013

Application Flow Cytometry
Fixation Formaldehyde
Sample Human Cell (BREAST CANCER TISSUE)
Specification BREAST CANCER TISSUE
Gating Strategy CD44 POSITIVE
Permeabilization Yes - BD FACS BUFFER
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Submitted Dec 09 2013

Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Human Cell lysate - whole cell (MCF7)
Specification MCF7
Total protein in input 100 µg
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Submitted Dec 09 2013

1-10 of 26 Abreviews or Q&A

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