Recombinant
RabMAb

Anti-SOX2 antibody [EPR3131] (ab92494)

Overview

  • Product name
    Anti-SOX2 antibody [EPR3131]
    See all SOX2 primary antibodies
  • Description
    Rabbit monoclonal [EPR3131] to SOX2
  • Tested applications
    Suitable for: WB, Sandwich ELISA, IHC-P, IHC - Wholemount, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Mouse, Human, Leucoraja erinacea
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human SOX2.

  • Positive control
    • WB: NCCIT, F9 and MCF-7 cell lysates. IHC-P: Human gliocytoma, breast carcinoma, fetal stomach, fetal lung and embryonal carcinoma tissues. ICC/IF: NCCIT cells.
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab92494 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 1/1000 - 1/2000. Detects a band of approximately 35 kDa (predicted molecular weight: 34 kDa).
Sandwich ELISA Use a concentration of 0.5 µg/ml.

For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Rabbit monoclonal [EPR3131] to SOX2 (ab92494) as Capture.

IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

For unpurified use at 1/60.

IHC - Wholemount Use at an assay dependent concentration.
ICC/IF 1/100.

For unpurified use at 1/60.

  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function
      Transcription factor that forms a trimeric complex with OCT4 on DNA and controls the expression of a number of genes involved in embryonic development such as YES1, FGF4, UTF1 and ZFP206 (By similarity). Critical for early embryogenesis and for embryonic stem cell pluripotency.
    • Involvement in disease
      Defects in SOX2 are the cause of microphthalmia syndromic type 3 (MCOPS3) [MIM:206900]. Microphthalmia is a clinically heterogeneous disorder of eye formation, ranging from small size of a single eye to complete bilateral absence of ocular tissues (anophthalmia). In many cases, microphthalmia/anophthalmia occurs in association with syndromes that include non-ocular abnormalities. MCOPS3 is characterized by the rare association of malformations including uni- or bilateral anophthalmia or microphthalmia, and esophageal atresia with trachoesophageal fistula.
    • Sequence similarities
      Contains 1 HMG box DNA-binding domain.
    • Post-translational
      modifications
      Sumoylation inhibits binding on DNA and negatively regulates the FGF4 transactivation.
    • Cellular localization
      Nucleus.
    • Information by UniProt
    • Database links
    • Alternative names
      • ANOP3 antibody
      • cb236 antibody
      • Delta EF2a antibody
      • lcc antibody
      • MCOPS3 antibody
      • MGC148683 antibody
      • MGC2413 antibody
      • RGD1565646 antibody
      • Sex determining region Y box 2 antibody
      • SOX 2 antibody
      • Sox2 antibody
      • SOX2_HUMAN antibody
      • SRY (sex determining region Y) box 2 antibody
      • SRY box containing gene 2 antibody
      • SRY related HMG box 2 antibody
      • SRY related HMG box gene 2 antibody
      • SRY-box 2 antibody
      • Transcription factor SOX 2 antibody
      • Transcription factor SOX-2 antibody
      • ysb antibody
      see all

    Anti-SOX2 antibody [EPR3131] images

    • Cell line: F9 (mouse embryonal carcinoma)

      Target AbID: Ab92494 anti-Sox2, Ab150077 AlexaFluor®488 Goat anti-Rabbit secondary

      Counterstain AbID: Ab7291 anti-Tubulin (Mouse mAb), Ab150120 AlexaFluor®594 Goat anti-Mouse secondary

      Fixative: 4% PFA

      Permeabilisation: 0.1% Triton-X

      Nuclear counter stain: DAPI

      Comments: Confocal image showing negative staining on F9 cells

      Target primary antibody dilution: 1:200

      Target secondary antibody dilution: 1:1000 (2ug/mL)

      Counterstain primary antibody dilution: 1:1000 (1ug/mL)

      Counterstain secondary antibody dilution: 1:1000 (2ug/mL)

      Negative control 1 primary antibody dilution: 1:200 (1ug/mL)  (Ab92494)

      Negative control 1 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150120)

      Negative control 2 primary antibody dilution: 1:1000 (1ug/mL) (Ab7291)

      Negative control 2 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150077)

    • Cell line: NCCIT (human pluripotent embryonal carcinoma)

      Target AbID: Ab92494 anti-Sox2, Ab150077 AlexaFluor®488 Goat anti-Rabbit secondary

      Counterstain AbID: Ab7291 anti-Tubulin (Mouse mAb), Ab150120 AlexaFluor®594 Goat anti-Mouse secondary

      Fixative: 4% PFA

      Permeabilisation: 0.1% Triton-X

      Nuclear counter stain: DAPI

      Comments: Confocal image showing negative staining on NCCIT cells

      Target primary antibody dilution: 1:200

      Target secondary antibody dilution: 1:1000 (2ug/mL)

      Counterstain primary antibody dilution: 1:1000 (1ug/mL)

      Counterstain secondary antibody dilution: 1:1000 (2ug/mL)

      Negative control 1 primary antibody dilution: 1:200 (1ug/mL) (Ab92494)

      Negative control 1 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150120)

      Negative control 2 primary antibody dilution: 1:1000 (1ug/mL) (Ab7291)

      Negative control 2 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150077)

    • Cell line: NIH/3T3 (mouse embryonic fibroblast cell line) (negative control).

      Target AbID: Ab92494 anti-Sox2, Ab150077 AlexaFluor®488 Goat anti-Rabbit secondary

      Counterstain AbID: Ab7291 anti-Tubulin (Mouse mAb), Ab150120 AlexaFluor®594 Goat anti-Mouse secondary

      Fixative: 4% PFA

      Permeabilisation: 0.1% Triton-X

      Nuclear counter stain: DAPI

      Comments: Confocal image showing negative staining on NIH/3T3 cells

      Target primary antibody dilution: 1:200

      Target secondary antibody dilution: 1:1000 (2ug/mL)

      Counterstain primary antibody dilution: 1:1000 (1ug/mL)

      Counterstain secondary antibody dilution: 1:1000 (2ug/mL)

      Negative control 1 primary antibody dilution: 1:200 (1ug/mL) (Ab92494)

      Negative control 1 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150120)

      Negative control 2 primary antibody dilution: 1:1000 (1ug/mL) (Ab7291)

      Negative control 2 secondary antibody dilution: 1:1000 (2ug/mL) (Ab150077)

    • All lanes : Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1000 dilution

      Lane 1 : F9 whole cell lysate
      Lane 2 : 4T1 whole cell lysate
      Lane 3 : Mouse hippocampus lysate
      Lane 4 : C6 whole cell lysate
      Lane 5 : Rat hippocampus lysate
      Lane 6 : Rat spinal cord lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa


      Exposure time : 3 minutes

      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • All lanes : Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1000 dilution

      Lane 1 : NCCIT whole cell lysate
      Lane 2 : PC-3 whole cell lysate
      Lane 3 : SK-OV-3 whole cell lysate
      Lane 4 : U2OS whole cell lysate
      Lane 5 : MCF7 whole cell lysate
      Lane 6 : HepG2 whole cell lysate
      Lane 7 : Human breast cancer tissue lysate
      Lane 8 : Human glioma lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa


      Exposure time : 3 minutes

      Blocking buffer: 5% NFDM/TBST
      Dilution buffer: 5% NFDM/TBST

    • Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1000 dilution (unpurified) + NCCIT cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Anti-SOX2 antibody [EPR3131] (ab92494) at 1/1500 dilution (purified) + F9 cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size : 34 kDa
      Observed band size : 34 kDa

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-SOX2 antibody [EPR3131] (ab92494) at 1/5000 dilution (unpurified)

      Lane 1 : NCCIT cell lysate
      Lane 2 : MCF-7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size : 34 kDa
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling SOX2 with unpurified ab92494 at 1/60. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gliocytoma tissue labelling SOX2 with purified ab92494 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling SOX2 with unpurified ab92494.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal stomach tissue labelling SOX2 with unpurified ab92494.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human fetal lung tissue labelling SOX2 with unpurified ab92494.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human embryonal carcinoma tissue labelling SOX2 with unpurified ab92494.

    • Immunohistochemistry (Formalin/PFA-fixed parffin-embedded sections) analysis of normal human lung tissue. Unpurified ab92494 shows negative staining.

    • Negative control: Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of negative human seminoma tissue using unpurified ab92494.

    • Immunocytochemsitry/Immunofluorescence analysis of NCCIT cells labelling SOX2 (green) with unpurified ab92494 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

    • Immunocytochemsitry/Immunofluorescence analysis of NCCIT cells labelling SOX2 (green) with purified ab92494 at 1/200. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

    • IHC - Wholemount analysis of Leucoraja erinacea embryo labelling SOX2 with unpurified ab92494 at 1/200. The sample was incubated with the primary antibody for 48 hours at 4°C in 10% fetal calf serum in PBT. Detection: DAB.

      See Abreview

    • IHC - Wholemount analysis of mouse blastocyst labelling SOX2 (pink) with unpurified ab92494 at 1/200. The sample was incubated with the primary antibody for 48 hours at 4°C. Nuclei stained with DAPI (grey).

      See Abreview

    • Standard Curve for SOX2 (Analyte: SOX2 protein (Human) (ab79950)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [57CT23.3.4] to SOX2 (ab75485) at 0.2µg/ml and Detector Antibody Rabbit monoclonal [EPR3131] to SOX2 (ab92494) at 0.5µg/ml.

    References for Anti-SOX2 antibody [EPR3131] (ab92494)

    This product has been referenced in:
    • Wei W  et al. Class I histone deacetylases are major histone decrotonylases: evidence for critical and broad function of histone crotonylation in transcription. Cell Res 27:898-915 (2017). WB ; Mouse . Read more (PubMed: 28497810) »
    • Bressan RB  et al. Efficient CRISPR/Cas9-assisted gene targeting enables rapid and precise genetic manipulation of mammalian neural stem cells. Development 144:635-648 (2017). ICC/IF . Read more (PubMed: 28096221) »

    See all 21 Publications for this product

    Product Wall

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Cow Tissue sections (Airway tissue)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA, pH 9.0
    Permeabilization
    No
    Specification
    Airway tissue
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 25°C
    Fixative
    Formaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Oct 21 2016

    Application
    Western blot
    Sample
    Pig Tissue lysate - whole (Heart)
    Gel Running Conditions
    Reduced Denaturing (4-12% bis tris gels)
    Loading amount
    100 µg
    Specification
    Heart
    Blocking step
    proprietary as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 24°C
    Username

    Miss. Laxmi Vemalapally

    Verified customer

    Submitted Dec 22 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Glioma)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA buffer, pH 9
    Permeabilization
    Yes - Triton 0.025%
    Specification
    Glioma
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Username

    Abcam user community

    Verified customer

    Submitted Jun 29 2015

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    • Endogenase peroxidase block with 3% H2O2 and DAKO S3022 antibody diluent (to reduce backgroun) as blocking agent for 5 minute(s) · Concentration: 100%
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Dako S1699
    Sample
    Human Tissue sections (patient brain tissues)
    Specification
    patient brain tissues
    Permeabilization
    No
    Fixative
    clinical samples (not sure)
    Username

    Abcam user community

    Verified customer

    Submitted Dec 04 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    IHC - Wholemount
    Sample
    Leucoraja erinacea Embryo (Whole embryo)
    Specification
    Whole embryo
    Username

    Dr. Andrew Gillis

    Verified customer

    Submitted Jul 24 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Glioblastoma primary cells)
    Loading amount
    30 µg
    Specification
    Glioblastoma primary cells
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Apr 16 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    IHC - Wholemount
    Sample
    Mouse Embryo (blastocyst)
    Specification
    blastocyst
    Username

    Abcam user community

    Verified customer

    Submitted Feb 06 2013

    Thank you for your inquiry.
    I received verification from Epitomics that they now recommend this antibody for use with mouse due to the fact that it produced a band at the expected molecular weight when tested in WB on a mouse cell line (F9).
    ...

    Read More

    Thank you for contacting us. John is away for a few weeks, my name is Keith.

    While I do not know exactly what you and John spoke of on the phone, from you email I have gathered that you are looking for very high affinity Sox2 antibodies for ...

    Read More

    Thank you for your reply.

    As ab95331 is a monoclonal antibody, it will only recognize and bind to one site on the protein, therefore, you would not be able to use it for both the capture and detection antibody. You would have to use it in conj...

    Read More

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