Anti-Estrogen Receptor beta antibody - ChIP Grade (ab3577)

Overview

  • Product nameAnti-Estrogen Receptor beta antibody - ChIP Grade
    See all Estrogen Receptor beta primary antibodies
  • Description
    Rabbit polyclonal to Estrogen Receptor beta - ChIP Grade
  • SpecificityBy Western blot, this antibody detects an ~55 kDa protein representing ER beta from Rat brain homogenate. Immunohistochemical staining of ER beta in Rat cerebellum results in cytoplasmic staining in Purkinje neurons. This antibody sufficiently supershifts Rat ER beta / ER response element (vitellogenin xenopus A2 gene).
  • Tested applicationsSuitable for: ICC/IF, EMSA, Gel supershift assays, IHC-P, IHC-Fr, WB, ICC, ChIP, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Hamster, Cow, Human, Pig, Non Human Primates, Baboon
  • Immunogen

    Synthetic peptide corresponding to Rat Estrogen Receptor beta aa 55-70.
    Sequence:

    AEPQKSPWCEARSLEH


    (Peptide available as ab3564)

  • Positive control
    • WB: Rat brain lysate IHC: Rat cerebellum, rat penis (see Jesmin et al), Ovine uterine artery endothelial cells (see Liao et al) ICC: Human coronary artery smooth muscle cells (see Han et al)

Properties

Applications

Our Abpromise guarantee covers the use of ab3577 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
ICC/IF Use a concentration of 0.5 µg/ml.
EMSA Use at an assay dependent concentration. PubMed: 18790053
Gel supershift assays Use a concentration of 1 - 5 µg/ml.
IHC-P Use a concentration of 1 - 2 µg/ml.
IHC-Fr Use a concentration of 1 - 2 µg/ml.
WB Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 59 kDa).Can be blocked with Human Estrogen Receptor beta peptide (ab3564).
ICC Use a concentration of 2 - 4 µg/ml.
ChIP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

Target

  • FunctionNuclear 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 specificityIsoform 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 similaritiesBelongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • DomainComposed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • Cellular localizationNucleus.
  • 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

Anti-Estrogen Receptor beta antibody - ChIP Grade images

  • ab3577 staining Estrogen Receptor beta in human placental sac tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, cut into 20 micron slices, blocked with 2.5% serum for 30 minutes at 25°C and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/200 in PBS) at 25°C for 90 minutes. An undiluted HRP-conjugated horse polyclonal was used as the secondary antibody.

    See Abreview

  • ab3577 labelling Estrogen Receptor beta by Immunocytochemistry/Immunofluorescence.

  • ab3577 at a concentration of 1-2µg/ml staining ER beta in primate amygdala.



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

    Image courtesy of an anonymous Abreview.

    Red - ab3577 at a 1/100 dilution
    Green - Actin loading control

    All samples loaded as 20µl.

    From left->right 1. Biorad ladder 2. B.end3 cells 20ug 3. B.end3 cells 40ug 4. B.end3 cells 60ug 5. B.end3 cells 80ug 6. Biorad ladder 7. Mouse Uterus Homogenate 10ug (+ Control) 8. Mouse Uterus Homogenate 20ug (+ Control) 9. Mouse Uterus Homogenate 40ug (+ Control) 10. Biorad Ladder

    The secondary used was an IRDye700DX conjugated rabbit polyclonal used at a 1/10000 dilution.

    See Abreview

References for Anti-Estrogen Receptor beta antibody - ChIP Grade (ab3577)

This product has been referenced in:
  • Persichetti P  et al. Oestrogen receptor-alpha and -beta expression in breast implant capsules: experimental findings and clinical correlates. J Plast Reconstr Aesthet Surg 67:308-15 (2014). IHC ; Human . Read more (PubMed: 24389289) »
  • Tam A  et al. Estradiol increases mucus synthesis in bronchial epithelial cells. PLoS One 9:e100633 (2014). WB, IHC-P ; Human . Read more (PubMed: 24964096) »

See all 17 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (placenta sac)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification placenta sac
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 26 2016

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (placenta)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification placenta
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 13 2015

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Horse Tissue sections (vagina)
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: 10 mM citrate, pH6.0
Permeabilization No
Specification vagina
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 05 2015

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.

P...

Read More

Thank you for your reply.

I am happy that the replacement antibody I sent you is producing better results in WB. It is not unusual for there to be observed differences in the molecular weight. This can be caused by differences in the buffer s...

Read More

Thank you for your inquiry.

Please follow this link to find the instruction on how to use the blocking peptide:

http://www.abcam.com/index.html?pageconfig=resource&rid=11378

I also attached the PDF to this email.
...

Read More

Thank you for your reply. The lysate can be boiled asyou would for your normal samples and loaded directly into the lane. I would recommend adding 4-6 uL of this lysate per lane. I hope this will give good results for you. If you have any additional qu...

Read More

Thank you so much for your reply and for sending all of this information.

These details will help us investigate the trouble with these antibodies. I apologize again for the trouble and for the inconvenience, and if there is anything else tha...

Read More

Thank you for your inquiry.

I found a publication that shows a WB were HepG2 cells are clearly negative for estrogen receptor beta:

Healter L. Rincalvage, Donald P. McDonnel and Cynthia M. Kuhn (2006). Expression of Functional Estro...

Read More

Thank you for contacting us.


I am sorry that this antibody did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by...

Read More

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"