RabMAb

Anti-Estrogen Receptor alpha抗体[E115] - ChIP Grade (ab32063)

概述

  • 产品名称Anti-Estrogen Receptor alpha抗体[E115] - ChIP Grade
    参阅全部 Estrogen Receptor alpha 一抗
  • 描述
    兔单克隆抗体[E115] to Estrogen Receptor alpha - ChIP Grade
  • 特异性Whilst this antibody showed positive staining against mouse, rat, and human tissue preparations by western blotting, the antibody failed to show positive IHC signal on mouse tissue sections when applied using our IHC testing conditions. For our in-house testing we tested the antibody on a mouse tissue array (spleen, lung, stomach, muscle, thymus, pancreas, liver, small intestine, brain, kidney).
  • 经测试应用适用于: ICC/IF, Flow Cyt, ChIP, WB, IHC-Pmore details
  • 种属反应性
    与反应: Mouse, Rat, Dog, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Estrogen Receptor alpha. The exact sequence is proprietary.
    (Peptide available as ab203371)

  • 阳性对照
    • WB: MCF-7 and T47-D cell lysates. IHC-P: Human breast carcinoma and endometrial carcinoma tissues. ICC/IF: MCF-7 cells. Flow Cyt: MCF-7 cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Alternative versions available:
    Anti-Estrogen Receptor alpha antibody (HRP) [E115] (ab194152)
    Anti-Estrogen Receptor alpha antibody (Alexa Fluor® 488) [E115] - (ab194150)
    Anti-Estrogen Receptor alpha antibody (Phycoerythrin) [E115] (ab209288)

    Anti-Estrogen Receptor alpha antibody (Low endotoxin, Azide free) [E115] (ab167390)

    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.

性能

应用

Our Abpromise guarantee covers the use of ab32063 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/200.
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

We recommend to use a 30 min blocking step (1XPBS/10%goat serum/0,3M Glycin).

ChIP Use 4 µg for 25 µg of chromatin.
WB 1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 67 kDa).Can be blocked with Estrogen Receptor alpha peptide (ab203371).
IHC-P 1/200.

For unpurified, use 1/50 - 1/100.

靶标

  • 功能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.
  • 序列相似性Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • 结构域Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • 翻译后修饰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).
  • 细胞定位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
  • 数据库链接
  • 别名
    • 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 [E115] - ChIP Grade 图像

  • Immunohistochemical analysis of human breast carcinoma using anti-Estrogen Receptor alpha (ab32063, unpurified) diluted 1:50

  • Immunofluorescent staining of MCF7 cells (fixed with 4% PFA and permeablized with TritonX 100) with purified ab32063 at a dilution of 1/250. An Alexa Fluor® 555 goat anti-rabbit antibody was used as the secondary at a dilution of 1/200. The panel on the right shows the DAPI counter-staining.

  • Chromatin was prepared from MCF-7+β-estraiol 30 min, and MCF-7 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 μg of chromatin, 4 μg of purified ab32063 (blue), and 20 μLl of anti-rabbit IgG sepharose beads. Rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR approach). Primers are located in the 2nd intron of TFF1 gene.

    MCF7 Cells were treated as below:

    MCF-7 starved overnight, then treated with 10 nM β-Estradiol in 2% FBS media for 30 min.

    Control MCF-7 was starved overnight, then in 2% FBS media for 30 mins.

    Primer information:

    Located to the 2 intron of TFF1 gene.

    Sequence:

    Forward: 5' -agtctcctccaacctgacctt-3'

    Reverse: 5' -ttccggccatctctcactat-3'

  • All lanes : Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) at 1/1000 dilution (purified)

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

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size : 67 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded human endometrial carcinoma with purified ab32063 at a working dilution of 1 in 200. The secondary antibody used is ab97051, a HRP goat anti-rabbit IgG (H+L), at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Estrogen Receptor alpha with purified ab32063 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei counterstained with DAPI (blue).

    Control 1: primary antibody (1/1000) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Overlay histogram showing MCF7 cells stained with unpurified ab32063 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab32063, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (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 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) at 1/500 dilution (unpurified) + MCF-7 cell lysate

    Predicted band size : 67 kDa
    Observed band size : 60 kDa (why is the actual band size different from the predicted?)
  • ChIP analysis using unpurified ab32063 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

Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)参考文献

This product has been referenced in:
  • Ruiz A  et al. Effect of hydroxychloroquine and characterization of autophagy in a mouse model of endometriosis. Cell Death Dis 7:e2059 (2016). IHC-P ; Mouse . Read more (PubMed: 26775710) »
  • Zhang W  et al. The correlation between DNMT1 and ERa expression and the methylation status of ERa, and its clinical significance in breast cancer. Oncol Lett 11:1995-2000 (2016). IHC-P ; Human . Read more (PubMed: 26998112) »

See all 19 Publications for this product

Product Wall

Abcam guarantees this product to work in the 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

提交于 Oct 17 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Feb 02 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
Username

Abcam user community

Verified customer

提交于 Jan 21 2016

Application IHC - Wholemount
Sample Mouse Tissue (Transplanted lung tumor section)
Specification Transplanted lung tumor section
Username

Abcam user community

Verified customer

提交于 Dec 21 2015

Application IHC - Wholemount
Sample Human Tissue (Invasive ductal carcinoma)
Specification Invasive ductal carcinoma
Username

Chi Li

Verified customer

提交于 Dec 19 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

提交于 Jul 13 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
Username

Abcam user community

Verified customer

提交于 Jun 26 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Horse Tissue sections (vagina)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification vagina
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

提交于 Jun 26 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (A549 lung cancer cells)
Permeabilization Yes - 0.2% Triton X-100
Specification A549 lung cancer cells
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 May 13 2015

1-10 of 28 Abreviews or Q&A

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