Name: Anti-Estrogen Receptor beta antibody [14C8]
SKU: ab288
Rating: 4 (9 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor beta antibody [14C8] (ab288)

Key features and details

Mouse monoclonal [14C8] to Estrogen Receptor beta
Suitable for: WB, IHC-P
Reacts with: Human
Isotype: IgG2b

Product name

Anti-Estrogen Receptor beta antibody [14C8]
See all Estrogen Receptor beta primary antibodies
Description

Mouse monoclonal [14C8] to Estrogen Receptor beta
Host species

Mouse

Tested Applications & Species

Application	Species
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment corresponding to Human Estrogen Receptor beta aa 1-153. Expressed in E.coli.
Database link: Q92731
Positive control
- WB: Recombinant fusion protein, human liver and intestine, HEK-293T cells. IHC-P: Endocervical epithelium, Lobular breast carcinoma and human breast carcinoma.
General notes

This product was changed from ascites to tissue culture supernatant on 23^rd October 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.40
Constituent: 100% PBS
Concentration information loading...
Purity

Immunogen affinity purified
Purification notes

Purified from TCS
Clonality

Monoclonal
Clone number

14C8
Myeloma

NS1
Isotype

IgG2b
Light chain type

kappa
Research areas

ChIP Related Products
- ChIP Kit (ab500)
Compatible Secondaries
- Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
- Goat Anti-Mouse IgG H&L (HRP) (ab205719)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control (ab170192)
Recombinant Protein
- Recombinant Human Estrogen Receptor beta protein (ab158385)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab288 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application	Species
IHC-P	Human
WB	Human

Application	Abreviews	Notes
WB	(2)	Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 59 kDa.
IHC-P	(5)	Use a concentration of 5 µg/ml.

Notes
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 59 kDa.
IHC-P Use a concentration of 5 µg/ml.

Function

Nuclear hormone receptor. Binds estrogens with an affinity similar to that of ESR1, and activates expression of reporter genes containing estrogen response elements (ERE) in an estrogen-dependent manner. Isoform beta-cx lacks ligand binding ability and has no or only very low ere binding activity resulting in the loss of ligand-dependent transactivation ability. DNA-binding by ESR1 and ESR2 is rapidly lost at 37 degrees Celsius in the absence of ligand while in the presence of 17 beta-estradiol and 4-hydroxy-tamoxifen loss in DNA-binding at elevated temperature is more gradual.
Tissue specificity

Isoform beta-1 is expressed in testis and ovary, and at a lower level in heart, brain, placenta, liver, skeletal muscle, spleen, thymus, prostate, colon, bone marrow, mammary gland and uterus. Also found in uterine bone, breast, and ovarian tumor cell lines, but not in colon and liver tumors. Isoform beta-2 is expressed in spleen, thymus, testis and ovary and at a lower level in skeletal muscle, prostate, colon, small intestine, leukocytes, bone marrow, mammary gland and uterus. Isoform beta-3 is found in testis. Isoform beta-4 is expressed in testis, and at a lower level in spleen, thymus, ovary, mammary gland and uterus. Isoform beta-5 is expressed in testis, placenta, skeletal muscle, spleen and leukocytes, and at a lower level in heart, lung, liver, kidney, pancreas, thymus, prostate, colon, small intestine, bone marrow, mammary gland and uterus. Not expressed in brain.
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.
Cellular localization

Nucleus.
Target information above from: UniProt accession Q92731 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2100 Human
- Omim: 601663 Human
- SwissProt: Q92731 Human
- Unigene: 660607 Human
- Unigene: 729020 Human
Alternative names
- ER BETA antibody
- ER-beta antibody
- Erb antibody
- ESR B antibody
- ESR BETA antibody
- ESR2 antibody
- ESR2_HUMAN antibody
- ESRB antibody
- ESTRB antibody
- estrogen nuclear receptor beta variant a antibody
- estrogen nuclear receptor beta variant b antibody
- estrogen receptor 2 (ER beta) antibody
- Estrogen receptor 2 antibody
- estrogen receptor beta 4 antibody
- Estrogen receptor beta antibody
- NR3A2 antibody
- Nuclear receptor subfamily 3 group A member 2 antibody
see all

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor beta antibody [14C8] (ab288)

Paraffin embedded human breast carcinoma tissue stained for Estrogen Receptor beta using ab288 at 1/200 dilution in immunohistochemical analysis.
Western blot - Anti-Estrogen Receptor beta antibody [14C8] (ab288)

All lanes : Anti-Estrogen Receptor beta antibody [14C8] (ab288) at 1/5000 dilution

Lane 1 : Non-transfected HEK-293T whole cell extracts
Lane 2 : Transfected HEK-293T whole cell extracts

Lysates/proteins at 30 µg per lane.

Predicted band size: 59 kDa

7.5% SDS-PAGE
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor beta antibody [14C8] (ab288)

ab288 staining Estrogen Receptor beta in infiltrating lobular carcinoma of the human breast tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde. Endogenous peroxidase blocking was with 3% H₂O₂ at room temperature for 30 minutes. Blocking with 1.5% goat serum (dilute goat serum by 1xPBS), at room temperature for 30min; antigen retrieval was by heat mediation in a citrate buffer, pH 6.0, 15min (Cuisinart Electric Pressure Cooker #EPC-1200, choose "high pressure"). Samples were incubated with primary antibody (1/100) for overnight at 4°C. An ABC HRP Kit Mouse IgG) was used as the secondary antibody (1/200), room temperature for 30min. Washed twice for 5 minutes using PBS. DAB was used as a substrate.
Western blot - Anti-Estrogen Receptor beta antibody [14C8] (ab288)

All lanes : Anti-Estrogen Receptor beta antibody [14C8] (ab288) at 1/500 dilution

Lane 1 : Human brain tissue lysate
Lane 2 : Human liver tissue lysate
Lane 3 : Human intestine tissue lysate

Lysates/proteins at 30 µg per lane.

Secondary
All lanes : Mouse IgG antibody (HRP) at 1/5000 dilution

Developed using the ECL technique.

Predicted band size: 59 kDa

7.5% gel.

Running condition: 80V, 15min; 140V, 40min.

Transfer condition: Semi-dry, 18 V, 60min (NC membrane)

blocking condition: 5% non-fat milk in TBST, RT, 60min.

primary antibody incubation: 4°C overnight.

Secondary antibody incubation: room temperature for 1 hour.

Washing conditions: 5 ml TBST, 4 x 5min.

ECL exposure.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor beta antibody [14C8] (ab288)

ab288 staining endocervical epithelium by IHC-P.

ChIP protocols
Immunohistochemistry protocols
Western blot protocols

Click here to view the general protocols

Publishing research using ab288? Please let us know so that we can cite the reference in this datasheet.

ab288 has been referenced in 79 publications.

Gu J et al. Targeting the ERß/Angiopoietin-2/Tie-2 signaling-mediated angiogenesis with the FDA-approved anti-estrogen Faslodex to increase the Sunitinib sensitivity in RCC. Cell Death Dis 11:367 (2020). PubMed: 32409702
Li H et al. CLPTM1L induces estrogen receptor ß signaling-mediated radioresistance in non-small cell lung cancer cells. Cell Commun Signal 18:152 (2020). PubMed: 32943060
Song P et al. Estrogen receptor ß inhibits breast cancer cells migration and invasion through CLDN6-mediated autophagy. J Exp Clin Cancer Res 38:354 (2019). PubMed: 31412908
Datta N et al. Promyelocytic Leukemia (PML) gene regulation: implication towards curbing oncogenesis. Cell Death Dis 10:656 (2019). PubMed: 31506431
López-Romero R et al. The expression of transcription factor BORIS and its association with the estrogen receptor beta (ER-ß) in cervical carcinogenesis. Int J Clin Exp Pathol 12:3208-3221 (2019). PubMed: 31934165

View all Publications for this product

Customer reviews and Q&As

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1-10 of 14 Abreviews or Q&A

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 - Buffer/Enzyme Used: Citrate pH6

Permeabilization

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

Abcam user community

Verified customer

Submitted Nov 28 2018

Application

Western blot

Sample

Human Cell lysate - whole cell (MDA-MB-231 cells)

Gel Running Conditions

Reduced Denaturing (4-20% Tris-Glycin gels)

Loading amount

50 µg

Specification

MDA-MB-231 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 28 2018

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (OV90)

Permeabilization

Yes - 0.25% Triton-X100 in PBS

Specification

OV90

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C

Fixative

Formaldehyde

Abcam user community

Verified customer

Submitted Oct 26 2016

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Blocking step

Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: 10mM sodium citrate, pH 6.0

Sample

Baboon Tissue sections (fetal ovary)

Specification

fetal ovary

Permeabilization

Fixative

Formaldehyde

Ms. Marcia Burch

Verified customer

Submitted Sep 25 2014

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Breast cancer)

Specification

Breast cancer

Fixative

Paraformaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: TRIS-EDTA-Buffer pH 9,0

Permeabilization

Yes - Wash Buffer with tween

Mr. Rudolf Jung

Verified customer

Submitted Dec 03 2012

Question

Dear sir, madam,

There is some literature stating that storage of tisse slides reduces antigen detection particulary for IHC and particulary for estrogen receptor. According to a recent article the main factor explaning the reduced staining intensity over time is the primary antibody used (Karlsson. Effects of long-term storage on the detection of proteins, DNA and mRNA in tissue microarray slides. J Histochem Cytochem 2011).

Do you have any experience about this topic related to Ab288? I just ordered two.

Thank you very much in advance.

Best regards,

Abcam community

Verified customer

Asked on Aug 15 2012

Answer

Thank you for your enquiry and your interest in our products.
The effect of formalin fixation and the long-term storage on the detectability of different proteins have been studies for some time. The longer the tissues are fixed and stored in formalin, less intensive staining can be obtained due to epitope accessibility and/or protein degradation. Generally, the optimal time for formalin fixation for most stains is around 3-7 days.
Currently, we do not have specific data/publications on ab288 regarding this issue.
If you need any further assistance in the future, please do not hesitate to contact me.

Abcam Scientific Support

Answered on Aug 15 2012

Question

LOT NUMBER GR60665-3 ORDER NUMBER 1004022 DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE whole cell lysate, human MCF-7, Keratinocyte PRIMARY ANTIBODY Concentration or dilution: 1/1000, 1/500, 1/200 Diluent buffer : 5% skim milk Incubation time : overnight Incubation temperature: 4℃ Wash Buffer : 1x TBST Number of washes : 3 times DETECTION METHOD ECL ANTIBODY STORAGE CONDITIONS -20C SAMPLE PREPARATION Lysis buffer : RIPA buffer Protease inhibitors: yes Phosphatase inhibitors Reducing agent : SDS and betaME Boiling for ≥5 min? yes AMOUNT OF PROTEIN LOADED 30 and 50ug/well ELECTROPHORESIS/GEL CONDITIONS 10% SDS-PAGE gel TRANSFER AND BLOCKING CONDITIONS Type of membrane : nitrocellulose Protein transfer verified: protein transfer was verified with ponceau S solution Blocking agent and concentration: 5% skim milk in 1xTBST Blocking time : 30min. Blocking temperature : room temperature SECONDARY ANTIBODY Concentration or dilution: 1/5000 Diluent buffer : 5% skim milk Incubation time : 2h Incubation temperature: room temperature Wash Buffer : 1x TBST Number of washes : 3 times HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? I tried to change the concentration of primary antibody and protein loading amount

Abcam community

Verified customer

Asked on Jan 09 2012

Answer

Thank you for taking time to liaise with this customer to complete our questionnaire. I am sorry to hear that this antibody is not providing satisfactory results. Having reviewed this case, I would like to offer some suggestions to help optimize the results from ab288. I would also appreciate if you can confirm some further details: 1.) The MCF-7 cells do express low levels of the estrogen receptor beta (http://www.ncbi.nlm.nih.gov/pubmed/10601611, https://www.abcam.com/Estrogen-Receptor-beta-antibody-14C8-ab288.html&tab=abreviews&intabreviewid=7400where it has been induced) and as far as I know, the expression in keratinocytes can also be variable. Does the customer know that the samples used do express high enough levels of Estrogen Receptor beta? If not, I can suggest to use another positive control, such as mouse brain extractor PC12 cell lysate for example. Does the customer have such asample? 2.) Secondly, I can also recommend touse BSA as a blocking buffer. Indeed, changing the blockingbuffercan significantly improve the results. Please seethe western blot image on the datasheet of ab9385 for an illustration. https://www.abcam.com/HRP-GAPDH-antibody-Loading-Control-ab9385.html (or use the following: https://www.abcam.com/HRP-GAPDH-antibody-Loading-Control-ab9385.html). 3.) I am not sure how the primary antibody incubation has been performed. I canhowever recommend to incubate the primary antibody overnight at 4 degrees Celsius. Should the suggestions not improve the results, please do let me know. In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund. I hope this information is helpful, and I thank you for your cooperation.

Abcam Scientific Support

Answered on Jan 09 2012

Application

Immunohistochemistry (Frozen sections)

Sample

Human Tissue sections (vaginal)

Specification

vaginal

Fixative

Paraformaldehyde

Permeabilization

Yes - 0.2% Triton in PBS

Blocking step

(agent) for 30 minute(s) · Concentration: 1% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Oct 17 2011

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (breast cancer)

Specification

breast cancer

Fixative

Paraformaldehyde

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: • Microwave slides in 10mM citrate buffer pH 6.0

Permeabilization

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Dr. W Xu

Verified customer

Submitted May 11 2010

Application

Western blot

Sample

Human Cell lysate - whole cell (MCF-7 cells)

Loading amount

50 µg

Specification

MCF-7 cells

Treatment

Doxycycline for 48 hrs

Blocking step

Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5%

Dr. George Skliris

Verified customer

Submitted May 09 2007

1-10 of 14 Abreviews or Q&A

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Key features and details

Overview

Product name

Description

Host species

Tested Applications & Species

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Purification notes

Clonality

Clone number

Myeloma

Isotype

Light chain type

Research areas

Associated products

ChIP Related Products

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Domain

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

References (79)

Customer reviews and Q&As

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor beta antibody [14C8]

Western blot abreview for Anti-Estrogen Receptor beta antibody [14C8]

Immunocytochemistry/ Immunofluorescence abreview for Anti-Estrogen Receptor beta antibody [14C8]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor beta antibody [14C8]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor beta antibody [14C8]

Immunohistochemistry (Frozen sections) abreview for Anti-Estrogen Receptor beta antibody [14C8]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Estrogen Receptor beta antibody [14C8]

Western blot abreview for Anti-Estrogen Receptor beta antibody [14C8]