Overview

  • Product name
  • Description
    Rabbit polyclonal to SOX9
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, Sandwich ELISA, IHC-Pmore details
    Unsuitable for: ICC/IF or IHC-Fr
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Cow, Dog, Common marmoset
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human SOX9.

    Read Abcam's proprietary immunogen policy (Peptide available as ab27845.)

  • Positive control
    • This antibody gave a positive signal in the following lysates: Neuroectoderm (HESCS differentiated) Day 5 and Day 10 - Whole Cell, Caco-2 Whole Cell, SW480 Whole Cell, Mouse Fetus (14 day old) Tissue

Properties

Applications

Our Abpromise guarantee covers the use of ab26414 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 Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 56 kDa).
Sandwich ELISA Use a concentration of 0.1 µg/ml. For sandwich ELISA, use this antibody as Detection at 0.1 µg/ml with Mouse monoclonal to SOX9 (ab58191) as Capture.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC/IF or IHC-Fr.
  • 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

    • Immunohistochemical analysis of formalin-fixed, paraffin embedded mouse spleen tissue, staining SOX9 with ab26414.

      Tissue was fixed with 10% Neutral Buffered Formalin and blocked with 1.5% serum for 45 minutes 21°C; antigen retrieval was by heat mediation in a Tris-EDTA buffer (pH 9). Samples were incubated with primary antibody (0.8 µg/ml in 0.3% Triton X-100 in PBS) for 1 hours at 21°C. An biotin-conjugated goat anti-rabbit polyclonal IgG (7 µg/ml) was used as the secondary antibody.

      A = Primary antibody
      B = No primary antibody

      See Abreview

    • All lanes : Anti-SOX9 antibody (ab26414) at 2 µg/ml

      Lane 1 : Neuroectoderm (differentiated from human embryonic stem cells) Day 5 - Whole Cell Lysate
      Lane 2 : Neuroectoderm (differentiated from human embryonic stem cells) Day 10 - Whole Cell Lysate
      Lane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
      Lane 4 : SW480 (Human colon adenocarcinoma cell line) Whole Cell Lysate
      Lane 5 : Mouse Fetus (14 Day Old) Tissue Lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 56 kDa
      Observed band size: 56 kDa
      Additional bands at: 100 kDa, 90 kDa. We are unsure as to the identity of these extra bands.



      Secondary antibody goat anti-rabbit HRP preadsorbed (IgG H&L; ab97080)

    • Standard Curve for SOX9 ; dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal to SOX9 (ab58191) at 5µg/ml and Detector Antibody Rabbit polyclonal to SOX9 (ab26414) at 0.1µg/ml.
    • IHC-P image of SOX9 (ab26414) on E11.5 mouse embryo sections. The sections were fixed in paraformaldehyde and underwent heat mediated antigen retrieval using Tris/EGTA (pH9). The sections were then blocked in 1% BSA solution for 30 mins at 20°C. In these images ab26414 can be seen staining sections showing spinal cord and developing cartilage (top row), developing cartilage in vertebrae (bottom row).

      See Abreview

    • Anti-SOX9 antibody (ab26414) at 1 µg/ml + Mouse Fetus (14 Day Old) Tissue Lysate at 10 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 56 kDa
      Observed band size: 56 kDa


      Exposure time: 90 seconds

    References

    This product has been referenced in:
    • Hong Y  et al. In vitro study on the role of SOX9 in trastuzumab resistance of adenocarcinoma of the esophagogastric junction. Exp Ther Med 15:3103-3107 (2018). Read more (PubMed: 29456713) »
    • Han S  et al. Dicam promotes proliferation and maturation of chondrocyte through Indian hedgehog signaling in primary cilia. Osteoarthritis Cartilage 26:945-953 (2018). WB . Read more (PubMed: 29702220) »
    See all 38 Publications for this product

    Customer reviews and Q&As

    1-10 of 28 Abreviews or Q&A

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (growth plate)
    Permeabilization
    Yes - Triton X
    Specification
    growth plate
    Blocking step
    Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde

    Dr. Tina Yuan

    Verified customer

    Submitted Jan 04 2018

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (SAOS cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Specification
    SAOS cells
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Oct 27 2016

    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (Skin)
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 22°C
    Fixative
    Paraformaldehyde

    Ahmar Aziz

    Verified customer

    Submitted Apr 05 2016

    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (Intestine)
    Specification
    Intestine
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Feb 12 2014

    Application
    Western blot
    Loading amount
    30000 cells
    Gel Running Conditions
    Reduced Denaturing (11)
    Sample
    Human Cell lysate - whole cell (MCF-7 cells (human breast adenocarcinoma cell line)
    Specification
    MCF-7 cells (human breast adenocarcinoma cell line
    Treatment
    0.5% oxygen for 16hrs
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Nov 05 2013

    Application
    Western blot
    Loading amount
    30000 cells
    Gel Running Conditions
    Reduced Denaturing (11)
    Sample
    Mouse Cell lysate - whole cell (Mouse embryonic fibroblast (MEF))
    Specification
    Mouse embryonic fibroblast (MEF)
    Treatment
    0.5% oxygen for 6hrs
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Nov 05 2013

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
    Sample
    Marmoset (common) Tissue sections (Neonatal testis)
    Specification
    Neonatal testis
    Permeabilization
    No
    Fixative
    Formaldehyde

    Dr. 煜欽 林

    Verified customer

    Submitted Oct 31 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Spleen)
    Specification
    Spleen
    Fixative
    10% neutral buffered formalin
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH 9.0 (20 min, 98*C)
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 21°C

    Abcam user community

    Verified customer

    Submitted Jan 24 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Articular Cartilage and Aortic Valve Tissue)
    Specification
    Articular Cartilage and Aortic Valve Tissue
    Fixative
    10% NBF
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 45 minute(s) · Concentration: 1% · Temperature: RT°C

    Abcam user community

    Verified customer

    Submitted Jan 24 2013

    Question



    1) Abcam product code ab 26414
    2) Abcam order reference number or product batch number CCE4329146
    3) Description of the problem

    For my research I want to identify Sertoli cells in rat testes. By achieving this I perform double staining (immunohistochemical) for Sox9. I ordered the anti-sox9 antibody ab26414, but after a couple of attemps I still do not have a positive result, I just see background staining. I will give a brief description of the protocol I used:
    - deparaffinezed with xylene and ethanol
    - clean up with hydrogen peroxide in methanol, wash in PBS
    - perform antigen retrieval with sodium citrate 3x5min 98 degrees, cooling down, wash PBS
    - 5% goat serum
    - 1:100 primary antibody, incubate overnight 4 degrees, wash PBS
    - 1:200 secondary goat biotinylated antibody, incubate, wash PBS
    - 1:1000 ABC, incubate, wash PBS
    - 1:200 DAB
    - stained with hematoxylin
    I have changed the dilution of the primary antibody to 1:20 and 1:200 -> no result.
    Unfortunately, I have not made pictures of the testis. Last week I used this antibody on kidney and liver. Both should be positive controls and again I had no result. I did make pictures and those added as attachements.
    I also diluted the primary antibody 200 times, but the results were similar > no result.
    4) Sample preparation:
    Species testis = rat, kidney = cat
    Type of sample: Fresh frozen sections, perfusion fixed frozen sections, PFA/formalin fixed paraffin embedded sections, cells in culture, other: formalin fixed paraffin
    Sample preparation
    Positive control
    Negative control
    5) Fixation step
    Yes/No
    If yes: Fixative agent and concentration
    Fixation time
    Fixation temperature
    6) Antigen retrieval method microwaving 3 x 5min in sodium citrate buffer pH 6
    7) Permeabilization method:
    Did you do a permeabilization step (details please) or add permeabilizing agent in any dilution buffers?
    Permeabilizing agent and concentration:
    8) Blocking agent (eg BSA, serum…): goat serum
    Concentration 5%
    Blocking time 30min
    Blocking temperature room temperature
    9) Endogenous peroxidases blocked? 3% hydrogen peroxide in methanol 30 min at room temperature
    Endogenous biotins blocked? 1:1000 ABC 60min at room temperature
    10) Primary antibody (If more than one was used, describe in “additional notes”) :
    Concentration or dilution Used several : 1:20, 1:100, 1:200
    Diluent buffer PBS/BSAc
    Incubation time overnight at 4 degrees
    11) Secondary antibody:
    Species: goat
    Reacts against: rabbit
    Concentration or dilution 1:200
    Diluent buffer PBS/BSAc
    Incubation time 60min at room temperature
    Fluorochrome or enzyme conjugate
    12) Washing after primary and secondary antibodies:
    Buffer PBS
    Number of washes after primary 6 times 5 min on shaker and after secondary 6 times 5 min on shaker
    13) Detection method 1:200 DAB and finally hematoxylin
    14) How many times have you run this staining? 5 times
    Do you obtain the same results every time? yes
    What steps have you altered to try and optimize the use of this antibody? Different dilutions
    Document attachment: Attaching images of your IHC is strongly recommended and can greatly speed up our investigation of your problem.
    Please send this questionnaire by e-mail by cutting and pasting the text in your e-mail (no attachments other than images) to technical@abcam.com.

    Read More
    Answer

    Thank you for your enquiry regarding ab26414 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody.

    Though you have kindly provided some details, it would be much appreciated if I could get some more information which would help me identify the source of the problem.

    1) Abcam order reference number or product batch number CCE4329146

    This is not batch neither order number. Please check your record again and give us the requested information. The batch number should be on the label of the tubes and the order number on the package you received in the box.

    I look forward to hearing from you soon.

    Read More

    1-10 of 28 Abreviews or Q&A

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

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