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

製品の概要

  • 製品名
    Anti-Estrogen Receptor alpha antibody [C-542] - ChIP Grade
    Estrogen Receptor alpha 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [C-542] to Estrogen Receptor alpha - ChIP Grade
  • 特異性
    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.
  • アプリケーション
    適用あり: ChIP, IP, ICC/IF, WB, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Hamster, Human
    交差が予測される動物種: Sheep, Horse, Cow, Chimpanzee
  • 免疫原

    Synthetic peptide:

    C-YYITGEAEGFPATV

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

  • 特記事項
    Storage below -20°C is not recommended.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能
    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.
  • 配列類似性
    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • ドメイン
    Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • 翻訳後修飾
    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).
  • 細胞内局在
    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
  • 参照データベース
  • 別名
    • 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 画像

  • 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

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

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

Product Wall

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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

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投稿 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

投稿 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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投稿 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

投稿 Dec 09 2013

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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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