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

Overview

  • Product name
    Anti-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
  • Specificity
    This 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 applications
    Suitable 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 notes
    Storage 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

  • 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

  • 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.

    See Abreview

  • Cross-linking (X-chIP) analysis of human MCF7 nuclear cell lysate labeling Estrogen Receptor alpha with ab66102 at concentration 4 µg/µg chromatin. Formaldehyde was used as the cross-linking agent (15 minutes). Semiquantitative PCR used for detection. Cells were incubated for 16 hours at 4°C with chIP dilution buffer.

    See Abreview

  • Anti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade (ab66102) at 3 µg/ml + Human MCF7 whole cell lysate at 100 µg

    Secondary
    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

    This image is courtesy of an anonymous abreview.

    Blocking step: 5% milk for 1 hour at 37°C.

    See Abreview

References

This product has been referenced in:
  • Maselli A  et al. Low expression of estrogen receptor ß in T lymphocytes and high serum levels of anti-estrogen receptor a antibodies impact disease activity in female patients with systemic lupus erythematosus. Biol Sex Differ 7:3 (2016). Flow Cyt, ICC/IF . Read more (PubMed: 26759713) »

See 1 Publication for this product

Customer reviews and Q&As

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
Username

Mr. Michael Milne

Verified customer

Submitted Jun 03 2014

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

Submitted Jun 05 2015

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%
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

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
Username

Abcam user community

Verified customer

Submitted Dec 09 2013

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