Overview

  • Product name

    Anti-Estrogen Receptor beta antibody
    See all Estrogen Receptor beta primary antibodies
  • Description

    Rabbit polyclonal to Estrogen Receptor beta
  • Host species

    Rabbit
  • Specificity

    By Western blot, this antibody detects an ~55 kDa protein representing ER beta from Rat brain homogenate. Immunocytochemical staining of ER beta in cells overexpressing the Human protein results in nuclear staining.
  • Tested applications

    Suitable for: IHC-P, IHC-Fr, ICC, WB, Inhibition Assay, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Sheep, Human, Pig, Non human primates
    Predicted to work with: Hamster
  • Immunogen

    Synthetic peptide corresponding to Rat Estrogen Receptor beta aa 467-485.
    Sequence:

    CSSTEDSKNKESSQNLQSQ


    (Peptide available as ab5018)

  • Positive control

    • WB: Rat brain lysate IHC: testis, efferent ductules, epididymis, and vas deferens (see Zhou et al), Rat ovary (see Sanchez-Criado)

Properties

Applications

Our Abpromise guarantee covers the use of ab3576 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
IHC-P 1/50. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr 1/50.
ICC 1/100 - 1/1000.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 59 kDa).
Inhibition Assay Use at an assay dependent concentration.
ICC/IF 1/100 - 1/1000.

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

  • Immunofluorescence analysis of frozen murine thoracic aorta, staining Estrogen Receptor beta with ab3576 at 1/50 dilution (green: not merged with nucleus, or purple: merged with nucleus). Nuclei were stained with DAPI (blue) and actin fibers were stained red.
  • Immunofluorescent analysis using Estrogen Receptor beta polyclonal antibody (ab3576) shows nuclear localization of Estrogen Receptor beta in MCF7 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% BSA for 15 minutes at room temperature. Cells were probed with a rabbit polyclonal antibody recognizing Estrogen Receptor beta (ab3576), at a dilution of 1:100 for 1 hour at room temperature. Cells were washed with PBS and incubated with DyLight 488 goat-anti-rabbit IgG secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.
  • All lanes : Anti-Estrogen Receptor beta antibody (ab3576) at 1/2000 dilution

    Lane 1 : CAL region of Human brain - Whole cell lysate
    Lane 2 : EC region of Human brain - Whole cell lysate
    Lane 3 : HIP region of Human brain - Whole cell lysate
    Lane 4 : IT region of Human brain - Whole cell lysate
    Lane 5 : MF region of Human brain - Whole cell lysate
    Lane 6 : SP region of Human brain - Whole cell lysate

    Secondary
    All lanes : HRP conjugated Donkey Anti-rabbit IgG

    Predicted band size: 59 kDa
    Observed band size: 55 kDa
    why is the actual band size different from the predicted?



    This image is courtesy of an Abreview submitted on 3 February 2006.

    See Abreview

  • Immunolocalization of ER beta from CHO cells overexpressing the Human protein, using ab3576 (1ug/ml).

References

This product has been referenced in:

  • Zhou F  et al. Raloxifene, a promising estrogen replacement, limits TDP-25 cell death by enhancing autophagy and suppressing apoptosis. Brain Res Bull 140:281-290 (2018). Read more (PubMed: 29807145) »
  • Wang J  et al. G-1 exhibit antidepressant effect, increase of hippocampal ERs expression and improve hippocampal redox status in aged female rats. Behav Brain Res N/A:N/A (2018). Read more (PubMed: 30041006) »
See all 44 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Answer

Thank you for your enquiry and interest in our products.

I can confirm that ab3576: Anti-Estrogen Receptor beta has been used in different dilutions in IHC (P).

There are two protocols which were used the same buffer but slightly different temperature and both were successfully applied with this antibody:

A) Paraffin-embedded sections were dewaxed and rehydrated through graded alcohol. Heat-mediated antigen retrieval was performed at 100 C in 1 liter 0.01 M citrate buffer (pH 6.0). Concentration of antibody used: 5 µg/ml.

B) After washing, the sections were transferred to plastic jars containing 0.01 M citrate buffer (pH 6.0) and boiled in a microwave oven for 10 min at 700 W. Samples were allowed to cool for 20 min before washing in PBS. 1:500 dilution in PBS + 2% BSA.

If you need any further assistance in the future, please do not hesitate to contact me.

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Question

Catalog number(s): 3576
Product(s): Rabbit polyclonal to Estrogen Receptor beta

-----------------------------------
Telephone section
Q. Did you contact Abcam's Scientific Support by telephone in reference to this complaint?
A. Yes
Q. Was it clear from the phone options which number you needed to press to get your issue resolved?
A. Yes

Protocol advice section
Q. Did Abcam's Scientific Support give you protocol advice?
A.Yes
Q. How did you feel about being given protocol advice?
A.I was not expecting to be given protocol advice, but was happy to receive this
Q. Did the product work successfully after following the advice?
A.No
Q. Why have you not tried the protocol advice? (Select all that apply.)
A.

Free of charge replacement section
Q. Did Abcam's Scientific Support give you a free of charge replacement?
A. Yes

Q. Did the replacement product work successfully?
A.Yes
Credit note / refund section
Q. Did Abcam's Scientific Support give you a credit note or a refund?
A.Yes

Q. Would you have preferred a different outcome from receiving a credit note or refund?
A.No
Overall satisfaction section
Q. How satisfied are you with how the complaint was handled?
A.Very satisfied
Q. Could we have done anything to better resolve the problem?
A.
Q. Would you mind if our scientific support team contacted you to better understand what happened and reach an agreeable solution?
A.No, I would not mind
Q. Please share with us any final thoughts you may have about Abcam, the complaints process, or our products.
A.Not this time. Thanks.

Read More
Answer

Thank you for your participation in Abcam's post complaint survey.

Please let me know if there is anything else I can do for you, or if you have questions to any of our products (e.g. primary antibodies, lysates, secondary antibodies, kits, controls, gels, proteins/peptides, slides, biochemicals, conjugation kits).

Read More
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Kidney)
Antigen retrieval step
Heat mediated
Permeabilization
No
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 09 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Monkey Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 09 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 09 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Breast
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 09 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% normal buffered formalin

Abcam user community

Verified customer

Submitted Oct 09 2019

Application
Flow Cytometry
Sample
Rat Cell (Lung tissue)
Permeabilization
Yes - eBiosciences FoxP3 intracellular staining buffer set
Gating Strategy
Leukocytes
Specification
Lung tissue
Preparation
Cell harvesting/tissue preparation method: Lungs were dissected and digested using Collagenase IV+DNase
Sample buffer: GKN+10% FCS
Fixation
eBiosciences FoxP3 intracellular staining buffer set

Abcam user community

Verified customer

Submitted Jan 12 2016

Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.
The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.
I would like to reassure you that ab3576 is tested and covered by our 6 month guarantee for use in WB and human, mouse and rat samples. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.
Reviewing this case, I would like to offer some suggestions to help optimise the results from ab3576.
1.) Antibodies will produce high background if they are used too concentrated. I suggest to further dilute the antibody.
2.) background is often generated by the secondary antibody and therefore I can recommend to rum a "no primary" control.
3.) Cell lines often produce artifact due to prolonged culturing. We have reports from customers using this antibody in tissue samples with good results. I suggest to run a positive control with a suitable tissue lysate.
4.) Since Estrogen Receptor beta is a nuclear protein, I can suggest to prepare nuclear lysates to clean up the blot.
I would also appreciate if you can confirm some further details:
1.) What lysis buffer was used?
2.) There are 8 isoforms of this protein. They range from 35kDa to 59kDa. Do you which one are expressed in the cell lines used?
I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

Read More

Answer

Thank you very much for you reply.

I'm sending a free of charge vial of ab3576 on the order ***, which we expect to arrive around August 24. Your purchasing agent will receive an email update if there are further delays to expect.

Please keep me updated about the results with this new antibody, and let me know if there is anything else that we can do for you. I look forward to hearing from you.

Have a nice day!

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1-10 of 22 Abreviews or Q&A

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