Overview

  • Product name
    Anti-SOX2 antibody [20G5]
    See all SOX2 primary antibodies
  • Description
    Mouse monoclonal [20G5] to SOX2
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, IP, WB, Flow Cyt, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
    Predicted to work with: Sheep, Chicken, Xenopus laevis, Zebrafish, Xenopus tropicalis
  • Immunogen

    Recombinant full length protein corresponding to Human SOX2 aa 1-317. Expressed in bacteria.
    Database link: 6657

  • Positive control
    • IHC-P: Human lung squamous carcinoma tissue. Mouse esophagus tissue. ICC/IF: H9 embryonic stem cells. HEL 11.4 induced IPS cells. IP: NCCIT whole cell lysate. WB: NCCIT and NTERRA cell lysate. Flow Cytometry: H9 embryonic stem cells. HEL 11.4 induced IPS cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab171380 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 1/20 - 1/200.
IP Use at 5 µg/mg of lysate.
WB 1/1000 - 1/2000. Predicted molecular weight: 34 kDa.
Flow Cyt 1/100.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

ICC/IF 1/200.

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

Images

  • Immunohistochemistry analysis of SOX2 showing staining in the nucleus of paraffin-treated human lung squamous carcinoma (right) compared with a negative control without primary antibody (left).

    To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with a SOX2 monoclonal antibody (ab171380) diluted by 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Immunofluorescence analysis of formaldehyde-fixed H9 embryonic stem cells, labeling SOX2 using ab171380 (left panel) at a 1/200 dilution overnight.

    DAPI was used to stain the cell nuclei (central panel). Slides were washed with PBS and incubated with a fluorescein-conjugated secondary antibody at a 1/100 dilution.

  • Western blot analysis on immunoprecipitation pellet from NCCIT (Human pluripotent embryonic carcinoma cell line) cells.

    The antigen-antibody complex was formed by incubating 500 µg of NCCIT whole cell lysate with 5 µg of ab171380 overnight at 4°C. The immune-complex was then captured on 50 µl Protein A/G Plus Agarose. Captured immune-complexes were then washes extensively and eluted sample with loading dye (lane 1) or 25 µg of NCCIT whole cells lysate (lane 2) were resolved on a SDS PAGE gel. An anti-mouse IgG-HRP secondary antibody at a 1/10,000 dilution was incubated for at least 1 hour. Chemiluminescent detection was perfomed.

  • All lanes : Anti-SOX2 antibody [20G5] (ab171380) at 1/500 dilution

    Lane 1 : NCCIT (Human pluripotent embryonic carcinoma cell line) cell lysate
    Lane 2 : NTERRA cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : mouse IgG-HRP at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 34 kDa

  • Flow cytometry analysis of H9 embryonic stem cells labeling SOX2 (blue histogram), using ab171380 at a 1/100 dilution, or a mouse IgG (green histogram) at a 1/100 dilution. A fluorescein-conjugated secondary antibody at a 1/200 dilution was used for the analysis.

  • Immunofluorescence analysis of formaldehyde-fixed HEL 11.4 induced IPS cells, labeling SOX2 using ab171380 (left panel) at a 1/200 dilution overnight. DAPI was used to stain the cell nuclei (central panel). Slides were washed with PBS and incubated with a fluorescein-conjugated secondary antibody at a 1/100 dilution.

  • Immunohistochemistry analysis of SOX2 showing staining in the nucleus of paraffin-treated mouse esophagus tissue (right) compared with a negative control without primary antibody (left).

    To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 minutes. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with a SOX2 monoclonal antibody (ab171380) diluted by 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

  • Flow cytometry analysis of HEL 11.4 induced IPS cells labeling SOX2 (blue histogram), using ab171380 at a 1/100 dilution, or a mouse IgG (green histogram) at a 1/100 dilution. A fluorescein-conjugated secondary antibody at a 1/200 dilution was used for the analysis.

References

This product has been referenced in:
  • Zeng H  et al. microRNA-129-5p suppresses Adriamycin resistance in breast cancer by targeting SOX2. Arch Biochem Biophys 651:52-60 (2018). WB . Read more (PubMed: 29802821) »
  • Than-Trong E  et al. Neural stem cell quiescence and stemness are molecularly distinct outputs of the Notch3 signalling cascade in the vertebrate adult brain. Development 145:N/A (2018). Read more (PubMed: 29695612) »
See all 9 Publications for this product

Customer reviews and Q&As

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

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Submitted Nov 28 2017

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