製品の概要

  • 製品名
    Anti-Estrogen Receptor beta antibody [14C8]
    Estrogen Receptor beta 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [14C8] to Estrogen Receptor beta
  • 由来種
    Mouse
  • 特異性
    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.
  • アプリケーション
    適用あり: WB, Flow Cyt, IHC-P, ICC/IF, IHC-Frmore details
  • 種交差性
    交差種: Mouse, Human, Monkey, Baboon
  • 免疫原

    Recombinant fragment corresponding to Human Estrogen Receptor beta aa 1-153. Expressed in E.coli.
    Database link: Q92731

  • ポジティブ・コントロール
    • WB: Recombinant fusion protein, human liver and intestine. IHC-P: endocervical epithelium, Lobular breast carcinoma.
  • 特記事項

    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.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
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.

ターゲット情報

  • 機能
    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.
  • 組織特異性
    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.
  • 配列類似性
    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • ドメイン
    Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • 細胞内局在
    Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • ab288 staining Estrogen Receptor beta in infiltrating lobular carcinoma of the 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.

  • 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. 

  • ab288 staining endocervical epithelium by IHC-P.
  • All lanes : Anti-Estrogen Receptor beta antibody [14C8] (ab288) at 1/5000 dilution

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

    Lysates/proteins at 30 µg per lane.

    Performed under non-reducing conditions.

    Predicted band size: 59 kDa

参考文献

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 63 Publications for this product

レビューと Q&A

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
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

Abcam user community

Verified customer

投稿 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

投稿 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

投稿 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
No
Fixative
Formaldehyde

Ms. Marcia Burch

Verified customer

投稿 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

投稿 Dec 03 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.

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
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

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

Dr. W Xu

Verified customer

投稿 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

投稿 May 09 2007

1-10 of 14 Abreviews or Q&A

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