Anti-Estrogen Receptor beta antibody [14C8] (ab288)

Reviews (9)Q&A (5)References (67)

Overview

  • 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

  • Specificity

    14C8 is specific for human ER-beta. The antibody has been tested in WB against recombinant protein. It has not yet been tested in our laboratory using any cell lysates and several publications suggest it can give weak bands in breast tumour extracts lysate. Strong Erb staining in epithelial cell nuclei. Occasional weak to moderate staining is seen in surrounding stromal and endothelial cell nuclei. Sporadic light cytoplasmic staining is sometimes observed. We have also successfully detected ERb in colon and ovarian tumors using the same antibody.

  • Tested applications

    Suitable for: WB, Flow Cyt, IHC-P, ICC/IF, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Human, Monkey, Baboon

  • 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. IHC-P: endocervical epithelium, Lobular breast carcinoma.

  • General notes

    This product was changed from ascites to tissue culture supernatant on 23rd 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.

Properties

Applications

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.

Application Abreviews Notes
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 59 kDa.
Flow Cyt 1/25.

ab170192 - Mouse monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

 
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Target

  • 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.
  • Information by UniProt

  • Database links

    • 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor beta antibody [14C8] (ab288)
      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)
      Western blot - Anti-Estrogen Receptor beta antibody [14C8] (ab288)
      All lanes : Anti-Estrogen Receptor beta antibody [14C8] (ab288) at 1/500 dilution

      All lanes : Human brain

      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)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Estrogen Receptor beta antibody [14C8] (ab288)
      ab288 staining endocervical epithelium by IHC-P.

    References

    This product has been referenced in:

    • Jiang R  et al. Serum Concentration of Genistein, Luteolin and Colorectal Cancer Prognosis. Nutrients 11:N/A (2019). Read more (PubMed: 30871032) »
    • Tang YQ  et al. Fructus Ligustri Lucidi modulates estrogen receptor expression with no uterotrophic effect in ovariectomized rats. BMC Complement Altern Med 18:118 (2018). Read more (PubMed: 29609586) »
    See all 67 Publications for this product

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

    Customer reviews and Q&As

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    1-10 of 14 Abreviews or Q&A

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

     Good
    Abreviews
    abreview image
    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
    No
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 28 2018

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

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 28 2018

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

     Inconclusive
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 26 2016

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

     Good
    Abreviews
    abreview image
    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
    No
    Fixative
    Formaldehyde
    Read More

    Ms. Marcia Burch

    Verified customer

    Submitted Sep 25 2014

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

     Good
    Abreviews
    abreview image
    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
    Read More

    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,

    Read More
    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.

    Read More

    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

    Read More
    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/index.html?datasheet=288&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/index.html?datasheet=9385 (or use the following: https://www.abcam.com/index.html?datasheet=9385). 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.

    Read More

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

     Good
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 17 2011

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

     Below Average
    Abreviews
    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
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Read More

    Dr. W Xu

    Verified customer

    Submitted May 11 2010

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

     Good
    Abreviews
    abreview image
    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%
    Read More

    Dr. George Skliris

    Verified customer

    Submitted May 09 2007

    1-10 of 14 Abreviews or Q&A

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