重组
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, 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)

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    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

图片

  • 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).

  • 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

  • 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.

  • 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'

  • 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.

  • 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

文献

This product has been referenced in:
  • Bi RY  et al. Estradiol upregulates voltage-gated sodium channel 1.7 in trigeminal ganglion contributing to hyperalgesia of inflamed TMJ. PLoS One 12:e0178589 (2017). IHC-Fr ; Rat . Read more (PubMed: 28582470) »
  • Ma F  et al. miR-148a Suppresses estrogen-induced viability and migration of breast cancer cells via inhibition of estrogen receptor a expression. Exp Ther Med 13:2515-2522 (2017). Read more (PubMed: 28565873) »

See all 25 Publications for this product

客户评价及客户问答

Application
Western blot
Sample
Mouse Tissue lysate - whole (liver)
Gel Running Conditions
Non-reduced Denaturing (12%)
Loading amount
30 µg
Specification
liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Nov 22 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (placenta)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
placenta
Blocking step
BSA as blocking agent for 30 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Dr. Kimitoshi Denda

Verified customer

提交于 Aug 31 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - nuclear (uterus)
Gel Running Conditions
Reduced Denaturing (10% NuPage MOPS)
Loading amount
10 µg
Treatment
ovariectomized, injected with estradiol for 1 hour
Specification
uterus
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Username

Sylvia Hewitt

Verified customer

提交于 Jun 27 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HUVEC)
Total protein in input
2000 µg
Immuno-precipitation step
Protein A
Specification
HUVEC
Username

Abcam user community

Verified customer

提交于 Mar 24 2017

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

提交于 Jan 09 2017

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

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

Dr. 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

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"

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