Name: Anti-SOX9 antibody [EPR14335-78]
SKU: ab185966
Rating: 5 (29 reviews)

Anti-SOX9 antibody [EPR14335-78] is a rabbit recombinant monoclonal antibody that is used to detect SOX9 in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Antibody clone EPR14335-78 is the most widely used clone for SOX9 on the market
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (AB185966), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Abcam Recommends

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IHC-P	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested
Mouse	Expected	Tested	Tested	Expected
Rat	Expected	Tested	Expected	Expected
Common marmoset	Predicted	Predicted	Predicted	Predicted
Cow	Predicted	Predicted	Predicted	Predicted
Dog	Predicted	Predicted	Predicted	Predicted
Pig	Predicted	Predicted	Predicted	Predicted

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000 - 1/10000	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Pig, Dog, Common marmoset, Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat mediated antigen retrieval with citrate buffer.
Species Rat	Dilution info 1/1000	Notes Perform heat mediated antigen retrieval with citrate buffer.
Species Human	Dilution info 1/1000	Notes Perform heat mediated antigen retrieval with citrate buffer.

Predicted
Predicted

Species	Dilution info	Notes
Species Pig, Dog, Common marmoset, Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1-5 µg/mL	Notes -
Species Human	Dilution info 1-5 µg/mL	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Pig, Dog, Common marmoset, Cow	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Pig, Dog, Common marmoset, Cow	Dilution info -	Notes -

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Target data

See full target information Transcription factor SOX-9

Function

Transcription factor that plays a key role in chondrocytes differentiation and skeletal development (PubMed:24038782). Specifically binds the 5'-ACAAAG-3' DNA motif present in enhancers and super-enhancers and promotes expression of genes important for chondrogenesis, including cartilage matrix protein-coding genes COL2A1, COL4A2, COL9A1, COL11A2 and ACAN, SOX5 and SOX6 (PubMed:8640233). Also binds to some promoter regions (By similarity). Plays a central role in successive steps of chondrocyte differentiation (By similarity). Absolutely required for precartilaginous condensation, the first step in chondrogenesis during which skeletal progenitors differentiate into prechondrocytes (By similarity). Together with SOX5 and SOX6, required for overt chondrogenesis when condensed prechondrocytes differentiate into early stage chondrocytes, the second step in chondrogenesis (By similarity). Later, required to direct hypertrophic maturation and block osteoblast differentiation of growth plate chondrocytes: maintains chondrocyte columnar proliferation, delays prehypertrophy and then prevents osteoblastic differentiation of chondrocytes by lowering beta-catenin (CTNNB1) signaling and RUNX2 expression (By similarity). Also required for chondrocyte hypertrophy, both indirectly, by keeping the lineage fate of chondrocytes, and directly, by remaining present in upper hypertrophic cells and transactivating COL10A1 along with MEF2C (By similarity). Low lipid levels are the main nutritional determinant for chondrogenic commitment of skeletal progenitor cells: when lipids levels are low, FOXO (FOXO1 and FOXO3) transcription factors promote expression of SOX9, which induces chondrogenic commitment and suppresses fatty acid oxidation (By similarity). Mechanistically, helps, but is not required, to remove epigenetic signatures of transcriptional repression and deposit active promoter and enhancer marks at chondrocyte-specific genes (By similarity). Acts in cooperation with the Hedgehog pathway-dependent GLI (GLI1 and GLI3) transcription factors (By similarity). In addition to cartilage development, also acts as a regulator of proliferation and differentiation in epithelial stem/progenitor cells: involved in the lung epithelium during branching morphogenesis, by balancing proliferation and differentiation and regulating the extracellular matrix (By similarity). Controls epithelial branching during kidney development (By similarity).

Alternative names

Transcription factor SOX-9, SOX9

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR14335-78
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-SOX9 antibody [EPR14335-78] (ab185966) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Anti-SOX9 antibody [EPR14335-78] (ab185966) was first used in a scientific publication in 2015 and has been cited over 245 times in peer reviewed journals. It's performance in IHC and Western Blot in mouse and human samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-SOX9 antibody [EPR14335-78] (ab185966) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-SOX9 antibody [EPR14335-78] (ab185966) has 26 independent reviews from customers.

Anti-SOX9 antibody [EPR14335-78] (ab185966) specifically detects SOX9 (UniProt ID: P48436; Molecular weight: 57kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR14335-78 - Anti-SOX9 antibody [EPR14335-78] - BSA and Azide free ab225541.

Antibody clone EPR14335-78 is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 647, Alexa Fluor^® 488, PE (Alexa Fluor® 647 Anti-SOX9 antibody [EPR14335-78] ab207677, Alexa Fluor® 488 Anti-SOX9 antibody [EPR14335-78] ab208427, PE Anti-SOX9 antibody [EPR14335-78] ab224019).

Top cited monoclonal for this target with >350 citations. Highly specific SOX9 antibody to study several important processes such as chondrogenesis and stem cell differentiation. SOX9 plays a multifaceted role in cancer development and progression. It functions as an oncogene in various cancers, including colorectal, ovarian and breast cancers. SOX9 promotes tumor initiation, proliferation, migration, invasion and chemoresistance by regulating key signalling pathways and gene expression. Additionally, SOX9 is involved in maintaining cancer stem cell properties, contributing to tumor growth and metastasis.

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

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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12 product images

This image is courtesy of a customer review submitted by Carl Hobbs, King's College London, United Kingdom
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
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.
Immunocytochemistry/ Immunofluorescence - Anti-SOX9 antibody [EPR14335-78] (ab185966)
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 Anti-alpha Tubulin antibody [DM1A] - Loading Control 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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and an anti-mouse AlexaFluor^® 594 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 (Intracellular) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Intracellular 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) (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 (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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Immunohistochemistry analysis of paraffin-embedded rat colon tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Immunohistochemistry analysis of paraffin-embedded mouse colon tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Immunohistochemistry analysis of paraffin-embedded human colon tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with hematoxylin.
This image is courtesy of a customer review submitted by Carl Hobbs, King's College London, United Kingdom
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX9 antibody [EPR14335-78] (ab185966)
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.
Immunocytochemistry/ Immunofluorescence - Anti-SOX9 antibody [EPR14335-78] (ab185966)
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.
Western blot - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Blocking and diluting buffer: 5% NFDM/TBST

SOX9 can be ubiquitinated or SUMOylated to higher molecular weight (PMID: 24155239, PMID: 16307912, PMID: 16554309, PMID: 32070068). Meanwhile, it has a truncated version as mini-Sox9 (PMID: 21297661，PMID: 27429045).

HeLa expresses very low level of SOX9 (PMID: 18296708, PMID: 18677406).
All lanes: Western blot - Anti-SOX9 antibody [EPR14335-78] (ab185966) at 1/1000 dilution
Lane 1: SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 4: Mouse colon tissue lysate at 20 µg
Lane 5: Mouse P0 bone tissue lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 75 kDa, 42 kDa, 56 kDa
Exposure time: 180s
Western blot - Anti-SOX9 antibody [EPR14335-78] (ab185966)
Western blot: Anti-Sox9 antibody [EPR14335-78] (ab185966) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab185966 was shown to bind specifically to Sox9. A band was observed at 50-70 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in Sox9 CRISPR-Cas9 edited cell line. The band observed in the CRISPR-Cas9 edited lysate lane below 50-70 kDa is likely to represent a truncated form of Sox9. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and Sox9 CRISPR-Cas9 edited HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-SOX9 antibody [EPR14335-78] (ab185966) at 1/1000 dilution
Lane 1: Wild-type HCT 116 cell lysate at 20 µg
Lane 2: Sox9 CRISPR-Cas9 edited HCT 116 cell lysate at 20 µg
Lane 3: SW480 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa, 70 kDa
Immunocytochemistry/ Immunofluorescence - Anti-SOX9 antibody [EPR14335-78] (ab185966)
SOX9 Immunocytochemistry/ Immunofluorescence staining of Primary hippocampal mouse neurons/glia DIV14. using rabbit Anti-SOX9 antibody
Immunofluorescence staining of SOX9 using ab185966 in primary hippocampal mouse neurons/glia (obtained from Transnetyx Tissue by BrainBits LLC cat.no. C57EHP) DIV14. The cells were fixed with 4% formaldehyde (10 min) permeabilized with 0.1% TritonX-100 (in PBS) for 5 mins 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 1 μg/ml, Anti-GFAP antibody - Astrocyte Marker ab4674 (anti-GFAP) at 1/1000 dilution and Anti-NeuN antibody [1B7] - Neuronal Marker ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution (shown in green) Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed ab150176 Goat Anti-Chicken IgY H&L (Alexa Fluor^® 594) preadsorbed (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) (shown in purple) all secondary antibodies at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).
As expected most GFAP positive cells are also SOX9 positive while NeuN positive cells are SOX9 negative. SOX9 positive cells which are not GFAP positive (e.g. asterisk) are likely neural stem cells/ oligodendrocyte precursor cells present in the culture.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.