Recombinant Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4097] to Estrogen Receptor alpha
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IHC-Fr, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-Estrogen Receptor alpha antibody [EPR4097]
See all Estrogen Receptor alpha primary antibodies -
Description
Rabbit monoclonal [EPR4097] to Estrogen Receptor alpha -
Host species
Rabbit -
Specificity
Expression levels of ER alpha protein vary with sample type. -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IHC-Fr, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: MCF7 and T47-D cell lysates. IHC-Fr: Frozen Human cervix and uterus tissue sections. IHC-P: Human breast ductal infiltrating carcinoma and normal breast tissues. ICC/IF: MCF-7 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
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
-
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR4097 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
- Anti-Estrogen Receptor alpha antibody [EPR4097] - BSA and Azide free (ab167610)
- Alexa Fluor® 647 Anti-Estrogen Receptor alpha antibody [EPR4097] (ab205851)
- Alexa Fluor® 594 Anti-Estrogen Receptor alpha antibody [EPR4097] (ab207046)
- Alexa Fluor® 555 Anti-Estrogen Receptor alpha antibody [EPR4097] (ab279333)
-
Assay kits
-
ChIP Related Products
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Positive Controls
-
Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab108398 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 |
---|---|---|
Flow Cyt (Intra) |
Use at an assay dependent concentration.
|
|
WB | (5) |
1/1000 - 1/10000. Predicted molecular weight: 66 kDa.
For unpurified use at 1/200 - 1/1000. |
IHC-P | (3) |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/30. |
IHC-Fr |
Use a concentration of 5 µg/ml.
|
|
ICC/IF |
1/250.
For unpurified use at 1/30. |
Notes |
---|
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
1/1000 - 1/10000. Predicted molecular weight: 66 kDa. For unpurified use at 1/200 - 1/1000. |
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/30. |
IHC-Fr
Use a concentration of 5 µg/ml. |
ICC/IF
1/250. For unpurified use at 1/30. |
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
modificationsPhosphorylated 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
- Entrez Gene: 2099 Human
- Omim: 133430 Human
- SwissProt: P03372 Human
- Unigene: 208124 Human
-
Alternative names
- 7*/654 isoform antibody
- 7*/819 2 isoform antibody
- 7*/822 isoform antibody
see all
Images
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue labelling Estrogen Receptor alpha with purified ab108398 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.
-
All lanes : Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398) at 1/2000 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
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: 66 kDa
Observed band size: 68 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsBlocking and diluting buffer: 5% NFDM/TBST
-
Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398) at 1/1000 dilution + GST-Tagged Recombinant Human Estrogen Receptor beta (aa 1 to 323) protein 100ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 66 kDa
Exposure time: 180 secondsBlocking/Diluting buffer and concentration: 5% NFDM/TBST
Lane 1: Recombinant Human Estrogen Receptor beta (aa 1 to 323) protein, (Cat#: ab158385)
There is no cross-reactivity between ab108398 and ER beta protein. -
All lanes : purified at 1/2000 dilution
Lane 1 : MCF-7 cell lysate
Lane 2 : T47-D cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 66 kDa
Observed band size: 67 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
-
Immunocytochemistry/ Immunofluorescence - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunocytochemsitry/Immunofluorescence analysis of MCF-7 cells labelling Estrogen Receptor alpha (green) with purified ab108398 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).
-
ab108398 staining Estrogen Receptor alphain the human cell line MCF-7 (human breast carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluorr® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast ductal infiltrating carcinoma tissue labelling Estrogen Receptor alpha with unpurified ab108398.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human normal tonsil tissue. Unpurified ab108398 shows negative staining.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human colonic adenocarcinoma tissue. Unpurified ab108398 shows negative staining.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human lung adenocarcinoma tissue. Unpurified ab108398 shows negative staining.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor alpha antibody [EPR4097] (ab108398)
Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of human cervical carcinoma tissue. Unpurified ab108398 shows negative staining.
-
IHC image of Estrogen Receptor alpha staining in a section of frozen human uterus* performed on a Leica Biosystems BOND® RX instrumen using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab108398, 5 ug/ml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
-
IHC image of Estrogen Receptor alpha staining in a section of frozen human cervix* performed on a Leica Biosystems BOND® RX instrumen using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab108398, 5 ug/ml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
-
Negative control image: IHC image of Estrogen Receptor alpha staining in a section of frozen human hippocampus* performed on a Leica Biosystems BOND® RX instrumen using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab108398, 5 ug/ml, 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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
Protocols
Datasheets and documents
-
SDS download
-
Datasheet download
References (25)
ab108398 has been referenced in 25 publications.
- Cheng JN et al. A Role for TGFß Signaling in Preclinical Osteolytic Estrogen Receptor-Positive Breast Cancer Bone Metastases Progression. Int J Mol Sci 22:N/A (2021). PubMed: 33923316
- Cheng R et al. Expression of IL17A in endometrial carcinoma and effects of IL17A on biological behaviour in Ishikawa cells. J Int Med Res 48:300060520950563 (2020). PubMed: 32878530
- Gao Y et al. Acetylation of histone H3K27 signals the transcriptional elongation for estrogen receptor alpha. Commun Biol 3:165 (2020). PubMed: 32265480
- Zhang KJ et al. miR-574-5p attenuates proliferation, migration and EMT in triple-negative breast cancer cells by targeting BCL11A and SOX2 to inhibit the SKIL/TAZ/CTGF axis. Int J Oncol 56:1240-1251 (2020). PubMed: 32319565
- Han Y et al. Inhibition of CDK2 reduces EZH2 phosphorylation and reactivates ERa expression in high-grade serous ovarian carcinoma. Am J Cancer Res 10:1194-1206 (2020). PubMed: 32368395