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RecombinantRabMAb

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

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  • SDS
Reviews (20)Q&A (11)References (118)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • ChIP - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Flow Cytometry (Intracellular) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • ChIP - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
  • Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E115] to Estrogen Receptor alpha - ChIP Grade
  • Suitable for: ICC/IF, ChIP, WB, Flow Cyt (Intra), IHC-P
  • Reacts with: Mouse, Rat, Human

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Alexa Fluor® 488 Carrier Free HRP PE

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Overview

  • Product name

    Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade
    See all Estrogen Receptor alpha primary antibodies
  • Description

    Rabbit monoclonal [E115] to Estrogen Receptor alpha - ChIP Grade
  • Host species

    Rabbit
  • Specificity

    Expression levels of ER alpha protein vary with sample type. Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) is unsuitable to test ovary and the tissues with low expression level of Estrogen Receptor alpha, such as kidney, lung and brain, in western blot. And it failed to show good IHC signal on mouse and rat tissue sections when using our IHC testing conditions. For our in-house testing we tested the antibody on a mouse tissue array (breast, spleen, lung, stomach, muscle, pancreas, liver, colon, brain, kidney).

  • Tested applications

    Suitable for: ICC/IF, ChIP, WB, Flow Cyt (Intra), IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab203371)

  • Positive control

    • WB: MCF-7 and T47-D cell lysates; Human uterus and ovary cancer tissue lysates; Mouse and rat uterus lysates; Rat and mouse pituitary whole tissue lysates. IHC-P: Human breast carcinoma and endometrial carcinoma tissues; human endometrium and breast tissues. ICC/IF: MCF-7 cells, 4T1 cells and GH3 cells. Flow Cyt (intra): MCF-7 cells. ChIP: Chromatin prepared from MCF-7+ß-estraiol 30 min cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E115
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Estrogen
    • Neuroscience
    • Endocrine system
    • Gonadotrophic axis
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Estrogen
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • Nuclear hormone receptors
    • Estrogen
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

Associated products

  • Alternative Versions

    • Anti-Estrogen Receptor alpha antibody [E115] - Low endotoxin, Azide free (ab167611)
    • Alexa Fluor® 488 Anti-Estrogen Receptor alpha antibody [E115] (ab194150)
    • HRP Anti-Estrogen Receptor alpha antibody [E115] (ab194152)
    • PE Anti-Estrogen Receptor alpha antibody [E115] (ab209288)
    • Anti-Estrogen Receptor alpha antibody [E115] - BSA and Azide free (ab271827)
  • Assay kits

    • Estrogen Receptor Transcription Factor Assay Kit (Colorimetric) (ab207203)
  • ChIP Related Products

    • ChIP Kit (ab500)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

    • Estrogen Receptor alpha peptide (ab203371)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human Estrogen Receptor alpha protein (ab82606)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
ICC/IF
1/200.
ChIP (1)
Use 4 µg for 25 µg of chromatin.
WB (5)
1/1000. Detects a band of approximately 60 kDa (predicted molecular weight: 67 kDa).Can be blocked with Estrogen Receptor alpha peptide (ab203371).
Flow Cyt (Intra)
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).

IHC-P (10)
1/200 - 1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

For unpurified, use 1/50 - 1/100. The antibody failed to show good IHC signal on mouse and rat tissue sections when applied using our IHC testing conditions.

Notes
ICC/IF
1/200.
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).
Flow Cyt (Intra)
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).

IHC-P
1/200 - 1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

For unpurified, use 1/50 - 1/100. The antibody failed to show good IHC signal on mouse and rat tissue sections when applied using our IHC testing conditions.

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.
  • Target information above from: UniProt accession P03372 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2099 Human
    • Entrez Gene: 13982 Mouse
    • Entrez Gene: 24890 Rat
    • Omim: 133430 Human
    • SwissProt: P03372 Human
    • SwissProt: P19785 Mouse
    • SwissProt: P06211 Rat
    • Unigene: 208124 Human
    • Unigene: 463262 Mouse
    • Unigene: 9213 Mouse
    • Unigene: 10595 Rat
    see all
  • Alternative names

    • 7*/654 isoform antibody
    • 7*/819 2 isoform antibody
    • 7*/822 isoform antibody
    • 8*/901 isoform antibody
    • 8*/941 isoform antibody
    • 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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    Immunohistochemical staining of paraffin embedded human endometrium tissue with ab32063 at a dilution of 1/5000. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP Polymer). The sample is counter-stained with hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0). 

    Nuclear staining on human endometrium.

  • Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    All lanes : Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063) at 1/200 dilution

    Lane 1 : Human uterus tissue lysates
    Lane 2 : Human kidney tissue lysates
    Lane 3 : Human brain tissue lysates
    Lane 4 : Mouse uterus tissue lysates
    Lane 5 : Mouse ovary tissue lysates
    Lane 6 : Mouse kidney tissue lysates
    Lane 7 : Mouse brain tissue lysates
    Lane 8 : Rat uterus tissue lysates
    Lane 9 : Rat ovary tissue lysates

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Predicted band size: 67 kDa
    Observed band size: 67 kDa


    Exposure time: 180 seconds


    Blocking and diluting buffer: 5% NFDM/TBST.

    The expression level of ER66 is high in uterus, moderate in ovary and low in kidney, lung, brain, liver, heart (PMID: 20861365, 24977106, 23675257, 23940668, 22070562), especially low in the tissues from male or young female animals (PMID: 26384003, 23940668). ab32063 is unsuitable to test ovary and the tissues with low expression level of Estrogen Receptor alpha in western blot.

  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 4T1 (mouse mammary gland carcinoma epithelial cell line) cells labelling Estrogen Receptor alpha with primary antibody anti-Estrogen Receptor alpha (ab32063) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing cytoplasmic and nuclear staining in 4T1 cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).

    The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2.0 µg/mL).

  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized GH3 (rat pituitary epithelial cell line) cells labelling Estrogen Receptor alpha with primary antibody anti-Estrogen Receptor alpha (ab32063) at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150081) secondary antibody at 1/1000 dilution (2.0 µg/mL). Confocal image showing cytoplasmic and nuclear staining in GH3 cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 dilution (2.5 µg/mL). The nuclear counter stain is DAPI (blue).

    The secondary antibody only control is : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (2.0 µg/mL).

  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    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).

  • ChIP - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    ChIP - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)This image is courtesy of an anonymous Abreview

    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

  • Flow Cytometry (Intracellular) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Flow Cytometry (Intracellular) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    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. 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    Immunohistochemical staining of paraffin embedded human breast carcinoma tissue with ab32063 at a dilution of 1/5000. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP Polymer). The sample is counter-stained with hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).

    Nuclear staining on human breast carcinoma.

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

    Lane 1 : MCF7 (Human breast adenocarcinoma epithelial cell). Whole cell lysates
    Lane 2 : T-47D (human mammary gland ductal carcinoma epithelial cell). Whole cell lysates
    Lane 3 : MDA-MB231 (Human breast adenocarcinoma epithelial cell) Whole cell lysates (Negative control)
    Lane 4 : HepG2 (Human hepatocellular carcinoma epithelial cell) Whole cell lysates (Negative control)
    Lane 5 : Human uterus whole tissue lysate
    Lane 6 : Human ovary whole tissue lysate
    Lane 7 : Human ovary cancer whole tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 67 kDa
    Observed band size: 68 kDa why is the actual band size different from the predicted?


    Exposure time: 50 seconds


    Blocking and diluting buffer: 5% NFDM/TBST. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    Immunohistochemical staining of paraffin embedded human breast tissue with ab32063 at a dilution of 1/5000. The secondary antibody used was Goat Anti-Rabbit IgG H&L (HRP Polymer). The sample is counter-stained with hematoxylin. Antigen retrieval was heat mediated using ab93684 (Tris/EDTA buffer, pH 9.0).

    Nuclear staining on human breast.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    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.

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

    Lane 1 : Rat pituitary whole tissue lysate
    Lane 2 : Mouse pituitary whole tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 67 kDa
    Observed band size: 68 kDa why is the actual band size different from the predicted?



    Exposure time: 1st lane: 85 seconds
                             2nd lane: 32 seconds

    Blocking and diluting buffer: 5% NFDM/TBST

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

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

  • Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    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.

  • Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    Western blot - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    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 - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
    ChIP - Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)

    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'

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

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

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References (118)

Publishing research using ab32063? Please let us know so that we can cite the reference in this datasheet.

ab32063 has been referenced in 118 publications.

  • Xiong Y  et al. Mixed-lineage leukaemia 1 contributes to endometrial stromal cells progesterone responsiveness during decidualization. J Cell Mol Med 25:297-308 (2021). PubMed: 33201593
  • Zhao Z  et al. Crosstalk between the muscular estrogen receptor a and BDNF/TrkB signaling alleviates metabolic syndrome via 7,8-dihydroxyflavone in female mice. Mol Metab 45:101149 (2021). PubMed: 33352311
  • Meng Q  et al. Estrogen prevent atherosclerosis by attenuating endothelial cell pyroptosis via activation of estrogen receptor a-mediated autophagy. J Adv Res 28:149-164 (2021). PubMed: 33364052
  • Wan X  et al. Estrogen Receptor a Mediates Doxorubicin Sensitivity in Breast Cancer Cells by Regulating E-Cadherin. Front Cell Dev Biol 9:583572 (2021). PubMed: 33614637
  • Shalabi MG  et al. The Prognostic Value of Estrogen Receptor ß Isoform With Correlation of Estrogen Receptor a Among Sudanese Breast Cancer Patients. Breast Cancer (Auckl) 15:1178223421998354 (2021). PubMed: 33716507
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1-10 of 20 Abreviews

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (placenta)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
placenta
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
Paraformaldehyde
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Mar 08 2021

Western blot abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Inconclusive
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Pancreas)
Gel Running Conditions
Reduced Denaturing (4-20%)
Loading amount
30 µg
Treatment
Tamoxifen 72 hours
Specification
Pancreas
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Dec 24 2020

Immunohistochemistry (Frozen sections) abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Inconclusive
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton X-100
Specification
Brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Mar 13 2020

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (MDA231 tumor taken out of Balb/C Nude mice)
Antigen retrieval step
Heat mediated
Permeabilization
Yes - Citrate pH6
Specification
MDA231 tumor taken out of Balb/C Nude mice
Blocking step
Innovex Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Nov 28 2018

Western blot abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (MD-MB-231 and MD-MB-468 cells)
Gel Running Conditions
Reduced Denaturing (4-20% Tris-Glycin gels)
Loading amount
50 µg
Specification
MD-MB-231 and MD-MB-468 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 28 2018

Western blot abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Nov 22 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Excellent
Abreviews
Abreviews
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. 公紀 伝田

Verified customer

Submitted Aug 31 2017

Western blot abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Good
Abreviews
Abreviews
abreview image
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
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The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Sylvia Hewitt

Verified customer

Submitted Jun 27 2017

Immunoprecipitation abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HUVEC)
Total protein in input
2000 µg
Immuno-precipitation step
Protein A
Specification
HUVEC
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Mar 24 2017

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 09 2017

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