Recombinant
RabMAb

Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396)

Overview

  • Product name
    Anti-Estrogen Receptor alpha (phospho S118) antibody [E91]
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Rabbit monoclonal [E91] to Estrogen Receptor alpha (phospho S118)
  • Specificity
    ab32396 only detects ER alpha phosphorylated on Serine 118.
  • Tested applications
    Suitable for: WB, ICC/IFmore details
    Unsuitable for: Flow Cyt or IHC-P
  • Species reactivity
    Reacts with: Human
  • Immunogen

    corresponding to Human Estrogen Receptor alpha.

  • Positive control
    • MCF7 cell lysate human breast adenocarcinoma
  • General notes

    A trial size is available to purchase for this antibody.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32396 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
WB 1/1000. Detects a band of approximately 66 kDa (predicted molecular weight: 66 kDa).
ICC/IF 1/200.

For unpurified use at 1/50 - 1/100 dilution. 

  • Application notes
    Is unsuitable for Flow Cyt or IHC-P.
  • 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

    • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma epithelial cell) treated with EGF (100ng/ml, 5min) and treated with Lambda Protein Phosphatase 31℃ for 2h cells labeling Estrogen receptor alpha (phospho S118) with purified ab32396 at 1:200 dilution (8.9μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    • All lanes : Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396) at 1/1000 dilution (purified)

      Lane 1 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysate
      Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor. Whole cell lysates
      Lane 3 : MCF-7 (Human breast adenocarcinoma epithelial cell) treated with epidermal growth factor. Whole cell lysates. Then the membrane was incubated with phosphatase.

      Lysates/proteins at 15 µg per lane.

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

      Predicted band size : 66 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)


      Exposure time : 5 seconds

      Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence analysis of MCF7 (Human breast adenocarcinoma cell line) cells labeling Estrogen Receptor alpha (phospho S118) with unpurified ab32396 at 5 μg/ml (1/200 dilution). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077, an AlexaFluor®488 Goat anti-Rabbit  was used as the secondary antibody at 2 μg/ml (1/1000 dilution). ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) was used to counterstain at 2.5 μg/ml (1/200 dilution). DAPI nuclear counterstain.
      Confocal image showing the signal increased after EGF (100ng/ml, 5 min) treatment and decreased after Lambda Protein Phosphatase treatment (31°C for 2 hours).

    • Immunohistochemical staining of (A) untreated and (B) Phosphatase-treated paraffin-embedded human breast adenocarcinoma using unpurified ab32396 at 1:50 dilution.

    • Dot blot analysis of Lane 1: Estrogen Receptor alpha (pS118) phospho peptide and Lane 2: Estrogen Receptor alpha non-phospho peptide labeling Estrogen Receptor alpha (phospho S118) with unpurified ab32396 at 1/1000 dilution. 5% NFDM/TBST was used as the diluting and blocking buffer. ab97051, Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time: 3 minutes.

    • Immunofluorescent staining of (A) untreated and (B) Phosphatase-treated MCF-7 cells using unpurified ab32396.

    • All lanes : Anti-Estrogen Receptor alpha (phospho S118) antibody [E91] (ab32396) at 1/1000 dilution (unpurified)

      Lane 1 : MCF7 cell lysate (untreated).
      Lane 2 : MCF7 cell lysate (treated with b-Estradiol and EGF).


      Predicted band size : 66 kDa
      Observed band size : 60 kDa (why is the actual band size different from the predicted?)

    References

    This product has been referenced in:
    • Ferraiuolo RM  et al. The cyclin-like protein, SPY1, regulates the ERa and ERK1/2 pathways promoting tamoxifen resistance. Oncotarget 8:23337-23352 (2017). WB ; Human . Read more (PubMed: 28423577) »
    • Yan X  et al. Ginsenoside Rd. promotes non-amyloidogenic pathway of amyloid precursor protein processing by regulating phosphorylation of estrogen receptor alpha. Life Sci N/A:N/A (2016). Read more (PubMed: 27825720) »

    See all 8 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    100 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (ovarian cancer cell lines and breast cancer cell l)
    Specification
    ovarian cancer cell lines and breast cancer cell l
    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

    Submitted Jul 23 2013

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (MCF7 and 293)
    Loading amount
    1e+006 cells
    Specification
    MCF7 and 293
    Treatment
    293 not treated, MCF7 ss and stimulated with E2
    Gel Running Conditions
    Reduced Denaturing (4-15% gradient gel)
    Blocking step
    Odyssey blocking buffer as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    Submitted Jul 26 2012

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

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