Anti-Estrogen Receptor alpha antibody [SP1] (ab16660)

Rabbit monoclonal Estrogen Receptor alpha antibody [SP1]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Human. Cited in 20 publication(s). Independently reviewed in 5 review(s).

Overview

  • Product name
    Anti-Estrogen Receptor alpha antibody [SP1]
    See all Estrogen Receptor alpha primary antibodies
  • Description
    Rabbit monoclonal [SP1] to Estrogen Receptor alpha
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IHC-FoFr, Flow Cytmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Pig
  • Immunogen

    Synthetic peptide within Human Estrogen Receptor alpha aa 550 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P03372

  • Epitope
    C-terminal
  • Positive control
    • WB: MCF7 cell lysate. IHC-P: Human breast carcinoma, cervix, breast, breast ductal carcinoma and ovarian adenocarcinoma tissue. ICC/IF: MCF7 cells. IHC-Fr: Mouse intestine tissue. FC: MCF7 cells
  • General notes

      

Properties

Applications

Our Abpromise guarantee covers the use of ab16660 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 1/25. Predicted molecular weight: 67 kDa.

Incubate for 1 hour at room temperature.

IHC-P 1/200.
ICC/IF 1/25 - 1/250.
IHC-FoFr Use at an assay dependent concentration. PubMed: 29403069
Flow Cyt 1/200.

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) cells labeling Estrogen Receptor alpha with purified ab16660 at 1/25 (8.5 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • IHC image of ab16660 staining Estrogen Receptor alpha in normal human cervix formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16660, 1/250 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Flow cytometry analysis of MCF7 (human breast adenocarcinoma epithelial cell) labeling Estrogen Receptor alpha with purified ab16660 at 1/200 dilution (1.06 µg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (black). Unlableled control - Unlabelled cells (blue).

  • Anti-Estrogen Receptor alpha antibody [SP1] (ab16660) at 1/25 dilution + lysate prepared from MCF7 cells

    Predicted band size: 67 kDa

  • ab16660 staining Estrogen Receptor alpha in MCF7 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab16660 at 1/250 dilution (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 594), at 2µg/ml (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunohistochemical analysis of paraformaldehyde-fixed, 0.1% Triton 0.05% Tween permeabilized frozen Mouse Tissue sections (Intestine) stained for Estrogen Receptor alpha (purple) using ab16660 at 1/100 dilution. Primary was incubated for 16 hours at 4°C with serum as blocking agent for 2 hours at 20°C.

    Secondary antibody was AffiniPure F(ab')2 Fragment Alexa Fluor® 647-conjugated Donkey anti-rabbit IgG at 1/500 dilution.

    HPRT-Cre/ERT2 x mT/mG mouse, treated with tamoxifen.

  • Formalin-fixed, paraffin-embedded human ovarian adenocarcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human breast ductal carcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human breast tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

  • Formalin-fixed, paraffin-embedded human breast carcinoma tissue stained for Estrogen Receptor alpha using ab16660 at 1/200 dilution in immunohistochemical analysis.

  • Human breast carcinoma stained with ab16660.

References

This product has been referenced in:
  • Koenigs MB  et al. Association of Estrogen Receptor Alpha Expression With Survival in Oropharyngeal Cancer Following Chemoradiation Therapy. J Natl Cancer Inst N/A:N/A (2019). IHC-P, IHC ; Human . Read more (PubMed: 30715409) »
  • Alzubi MA  et al. Separation of breast cancer and organ microenvironment transcriptomes in metastases. Breast Cancer Res 21:36 (2019). IHC-P, IHC ; Human . Read more (PubMed: 30841919) »
See all 29 Publications for this product

Customer reviews and Q&As

1-2 of 2 Q&A

Question
Answer

Thank you for your telephone enquiry this morning.


As discussed on the telephone, I can confirm these antibodies are different and will be detecting different eptiopes. They have also been tested ina slightly different list of applications and species.One other major difference is that ab16660is sold as tissue culture supernatant, ab27614 is purified.

During our discussions, you mentioned these may be used for diagnostics. Please notethat regrettablyour products are not accredited for clinical diagnostic use.However, we canprovide our 6 month guarantee for the applications and species listed on the individual datasheets.

We do sell a large range of IHC reagents now which I hope will be helpful to you. They can be found by searching on our website, or you may find it helpful to click on the links from the following IHC reagentspage:

https://www.abcam.com/index.html?pageconfig=resource&rid=12853

I hope this information is helpful. Should you have any further questions, please do not hesitate to contact us.

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Answer

Thank you for your enquiry. I am pleased to provide the following suggested positive controls for these antibodies. I have mainly chosen tissue that has been tested succesfully and shown in an image on the datasheets: 1.Anti-ErbB 2 antibody (ab2428) Human breast cancer 2.Anti-Progesterone Receptor antibody [SP2] (ab16661) Human breast carcinoma 3.Anti-Estrogen Receptor antibody [SP1] (ab16660) Human breast carcinoma 4.Anti-Myeloperoxidase antibody (ab45977) Human spleen tissue 5.Anti-TdT antibody (ab85148) Human testis 6.Anti-TMEM16A antibody (ab53212) Human liver 7.Anti-NSE antibody [EPR3377] (ab79757) Human brain tissue 8.Anti-CD23 antibody [EPR3617(IHC)] (ab92495) Human tonsil 9.Anti-CD10 antibody [56C6] (ab951) Lymph node tissue or human breast microepithelium 10.Anti-Melanoma antibody [HMB45 + DT101 + BC199] (ab732) Murine melanoma metastasis tissue 11.Anti-CD79a antibody [HM47/A9] (ab3121) Human Lymphoid tissue, tonsil 12.Anti-CD45RO antibody [UCHL-1] (ab86080) Human Tonsil, Thymus 13.Anti-CD45 antibody (ab10559) Human Tonsil tissue 14.Anti-CD30 antibody [CON6D/ C2] - BSA and Azide free (ab76518) Human Hodgekins lymphoma tissue 15.Anti-CD15 antibody [(Workshop IV) (BRA-4F1) our designation 238)] (ab115993) Human Hodgekins lymphoma tissue Human spleen I am providing a list ofslides that we sell which I hope will be helpful to you. I can recommend to review the online datasheet for more details. Human brain: ab4305Brain (human): hippocampus normal tissue slides https://www.abcam.com/index.html?datasheet=4305 (or use the following: https://www.abcam.com/index.html?datasheet=4305). Human lymph node: ab4350Lymph node (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4350 (or use the following: https://www.abcam.com/index.html?datasheet=4350). Human testis: ab4373Testis (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4373 (or use the following: https://www.abcam.com/index.html?datasheet=4373). Human liver: ab4348Liver (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4348 (or use the following: https://www.abcam.com/index.html?datasheet=4348). Human spleen: ab4372Spleen (human) normal tissue slides IHC-P, ISH https://www.abcam.com/index.html?datasheet=4372 (or use the following: https://www.abcam.com/index.html?datasheet=4372). I am sorry we do not have slides forthe following: Human tonsil Hodgekins lymphoma tissue Mouse melenoma tissue I hope this will be helpful to you. If you have any further questions,please do not hesitate to contact us.

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