Overview

  • Product name
    Anti-SOX9 antibody [EPR14335-78]
    See all SOX9 primary antibodies
  • Description
    Rabbit monoclonal [EPR14335-78] to SOX9
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
  • Immunogen

    Recombinant fragment within Human SOX9 aa 150-300 (internal sequence). The exact sequence is proprietary.
    Database link: P48436

  • Positive control
    • IHC-P: Mouse eye tissue. Rat, mouse and human colon tissue. Human breast carcinoma tissue. WB: SW480 and PC-3 cell lysate. ICC/IF: SW480, F9 and PC-3 cells. Flow Cyt: PC-3 cells.
  • General notes

      

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    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 ab185966 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
Flow Cyt Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 5 µg/ml.
WB 1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 56 kDa).
IHC-P 1/1000.

Target

  • Function
    Plays an important role in the normal skeletal development. May regulate the expression of other genes involved in chondrogenesis by acting as a transcription factor for these genes.
  • Involvement in disease
    Defects in SOX9 are the cause of campomelic dysplasia (CMD1) [MIM:114290]. CMD1 is a rare, often lethal, dominantly inherited, congenital osteochondrodysplasia, associated with male-to-female autosomal sex reversal in two-thirds of the affected karyotypic males. A disease of the newborn characterized by congenital bowing and angulation of long bones, unusually small scapulae, deformed pelvis and spine and a missing pair of ribs. Craniofacial defects such as cleft palate, micrognatia, flat face and hypertelorism are common. Various defects of the ear are often evident, affecting the cochlea, malleus incus, stapes and tympanum. Most patients die soon after birth due to respiratory distress which has been attributed to hypoplasia of the tracheobronchial cartilage and small thoracic cage.
  • Sequence similarities
    Contains 1 HMG box DNA-binding domain.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • campomelic dysplasia autosomal sex reversal antibody
    • CMD 1 antibody
    • CMD1 antibody
    • CMPD 1 antibody
    • CMPD1 antibody
    • SOX 9 antibody
    • Sox9 antibody
    • SOX9_HUMAN antibody
    • SRA 1 antibody
    • SRA1 antibody
    • SRXX2 antibody
    • SRXY10 antibody
    • SRY (sex determining region Y) box 9 (campomelic dysplasia autosomal antibody
    • SRY (sex determining region Y) box 9 antibody
    • SRY (sex determining region Y)-box 9 antibody
    • SRY (sex-determining region Y)-box 9 protein antibody
    • SRY related HMG box gene 9 antibody
    • Transcription factor SOX 9 antibody
    • Transcription factor SOX-9 antibody
    • transcription factor SOX9 antibody
    see all

Images

  • ab185966 staining SOX9 in developing eye of mouse tissue section by Immunohistochemistry (PFA perfusion fixed frozen sections).

    Tissue samples were fixed by perfusion with formaldehyde, cut into 20 micron slices, blocked with 2% BSA for 10 minutes at 21°C and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/1000 in PBS) at 21°C for 4 hours. A biotin-conjugated goat anti-rabbit polyclonal (1/300) was used as the secondary antibody.

    See Abreview

  • Anti-SOX9 antibody [EPR14335-78] (ab185966) at 1/20000 dilution + SW480 (Human colorectal adenocarcinoma cell line) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa

  • ab185966 staining Sox9 in F9 (Mouse embryonic testicular cancer cell line) cells.

    The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab185966 at a 5 μg/ml concentration and ab7291, mouse monoclonal [DM1A] to alpha Tubulin, at  1μg/ml concentration, followed by a further incubation at room temperature for 1 h with an anti-rabbit AlexaFluor® 488 (ab150081) at 2 μg/ml (shown in green) and an anti-mouse AlexaFluor® 594 (ab150120) at 2 μg/ml (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Flow Cytometry analysis of PC-3 (human prostate adenocarcinoma) cells labeling SOX9 with purified ab185966 at 1/120 (red).

    Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Anti-SOX9 antibody [EPR14335-78] (ab185966) at 1/5000 dilution + PC-3 (Human prostate adenocarcinoma cell line) cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 56 kDa

  • Immunohistochemistry analysis of paraffin-embedded rat colon tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.

  • Immunofluorescence analysis of 4% paraformaldehyde fixed PC-3 (Human prostate adenocarcinoma cell line) cells labeling SOX9 with ab185966 at 1/250 dilution (Left panel, red).

    Goat anti Rabbit IgG (Alexa Fluor®555) used as secondary antibody at 1/200 dilution. DAPI staining (Right panel, blue).

  • Immunohistochemistry analysis of paraffin-embedded mouse colon tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.

  • Immunohistochemistry analysis of paraffin-embedded human colon tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.

  • Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.

  • Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded pig small intestine sections labeling SOX9 with ab185966 at dilution of 1/2000.

    The secondary antibody used was a polyclonal goat anti-rabbit biotin conjugated antibody at a dilution of 1/300. The sample was counterstained with hematoxylin. Antigen retrieval was heat mediated using citric acid.

    The image shows intense enterocyte/goblet cell nuclear positivity, confined to the crypts of Lieberkühn.

    See Abreview

  • Immunofluorescence analysis of 4% paraformaldehyde fixed SW480 (Human colorectal adenocarcinoma cell line) cells labeling SOX9 with ab185966 at 1/250 dilution. Goat anti Rabbit IgG (Alexa Fluor®555) used as secondary antibody at 1/200 dilution. DAPI staining shown in blue.

References

This product has been referenced in:
See all 18 Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application
Immunohistochemistry (Frozen sections)
Sample
Cat Tissue sections (Optic nerve)
Permeabilization
Yes - Tween20
Specification
Optic nerve
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 12 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Monodelphis domestica Cell (cortex)
Permeabilization
Yes - 0,1% Triton X-100 in PBS
Specification
cortex
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5µg/mL · Temperature: 22°C
Fixative
Paraformaldehyde

Jelena Ban

Verified customer

Submitted Mar 22 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (PC3 Prostate cancer cells)
Permeabilization
Yes - 0.1% Triton X-100
Specification
PC3 Prostate cancer cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Dimitra Kalamida

Verified customer

Submitted Mar 22 2019

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Monodelphis domestica Tissue sections (spinal cord)
Antigen retrieval step
None
Permeabilization
Yes - 0.3% Triton X-100 in PBS for 1 hour
Specification
spinal cord
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
Paraformaldehyde

Jelena Ban

Verified customer

Submitted Nov 08 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Skin)
Permeabilization
No
Specification
Skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.25% · Temperature: 25°C
Fixative
Paraformaldehyde

Mr. Elena Ezhkova

Verified customer

Submitted Jan 09 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (mouse intestinal organoids)
Specification
mouse intestinal organoids
Blocking step
Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde

Sylke Hoehnel

Verified customer

Submitted Jun 20 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (brain)
Specification
brain
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C
Fixative
Paraformaldehyde

Wei Mo

Verified customer

Submitted Dec 22 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Colon
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 06 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Developing eye ( e16))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Developing eye ( e16)
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 06 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Developing lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Developing lung
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Sep 06 2016

1-10 of 11 Abreviews

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