Recombinant Anti-Estrogen Receptor alpha antibody [E115] - Low endotoxin, Azide free (ab167611)
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 - Low endotoxin, Azide free
- Suitable for: WB, ChIP, IHC-P, Flow Cyt (Intra), ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Estrogen Receptor alpha antibody [E115] - Low endotoxin, Azide free
See all Estrogen Receptor alpha primary antibodies -
Description
Rabbit monoclonal [E115] to Estrogen Receptor alpha - Low endotoxin, Azide free -
Host species
Rabbit -
Specificity
Expression levels of ER alpha protein vary with sample type. This antibody 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).
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Tested applications
Suitable for: WB, ChIP, IHC-P, Flow Cyt (Intra), ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- ICC/IF: MCF-7 cells, 4T1 cells and GH3 cells.
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General notes
ab167611 is the carrier-free version of ab32063.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
E115 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- 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)
- APC Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab310852)
- Alexa Fluor® 647 Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab311062)
- Alexa Fluor® 594 Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab311647)
- Alexa Fluor® 568 Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab312920)
- Alexa Fluor® 555 Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab313132)
- Anti-Estrogen Receptor alpha antibody [E115] - ChIP Grade (ab32063)
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab167611 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 |
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WB |
Use at an assay dependent concentration.
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ChIP |
Use at an assay dependent concentration.
Please refer to the original abID, ab32063, for information on recommended dilutions. |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Please refer to the original abID, ab32063, for information on recommended dilutions. |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
Please refer to the original abID, ab32063, for information on recommended dilutions. |
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ICC/IF |
Use at an assay dependent concentration.
Please refer to the original abID, ab32063, for information on recommended dilutions. |
Notes |
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WB
Use at an assay dependent concentration. |
ChIP
Use at an assay dependent concentration. Please refer to the original abID, ab32063, for information on recommended dilutions. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Please refer to the original abID, ab32063, for information on recommended dilutions. |
Flow Cyt (Intra)
Use at an assay dependent concentration. Please refer to the original abID, ab32063, for information on recommended dilutions. |
ICC/IF
Use at an assay dependent concentration. Please refer to the original abID, ab32063, for information on recommended dilutions. |
Target
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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
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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
see all -
Alternative names
- 7*/654 isoform antibody
- 7*/819 2 isoform antibody
- 7*/822 isoform antibody
see all
Images
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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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'
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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Clone E115 (ab167611) has been successfully conjugated by Abcam. This image was generated using Anti-Estrogen Receptor alpha antibody [E115] (PE). Please refer to ab209288 for protocol details.
ab209288 staining Estrogen Receptor alpha in T47D cells. The cells were fixed with 4% formaldehyde (10 min), 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 ab209288 at 1/500 dilution (Pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Clone E115 (ab167611) has been successfully conjugated by Abcam. This image was generated using Anti-Estrogen Receptor alpha antibody [E115] (Alexa Fluor® 488). Please refer to ab194150 for protocol details.
Overlay histogram showing T47D cells stained with ab194150 (red line). The cells were fixed with 4% formaldehyde (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 (ab194150, 1/50 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 488 (ab199091) used at the same concentration and conditions as the primary antibody. 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 T47D cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32063).
Protocols
Datasheets and documents
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Datasheet download
References (1)
ab167611 has been referenced in 1 publication.
- Huang S et al. Clinicopathological and Prognostic Significance of Klotho and Estrogen Receptors Expression in Human Hepatocellular Carcinoma. Turk J Gastroenterol 32:828-836 (2021). PubMed: 34787087