Recombinant Anti-SOX9 antibody [EPR14335] - BSA and Azide free (ab220812)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14335] to SOX9 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SOX9 antibody [EPR14335] - BSA and Azide free
See all SOX9 primary antibodies -
Description
Rabbit monoclonal [EPR14335] to SOX9 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Cow, Dog, Pig, Common marmoset -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: SW480, HeLa and NIH/3T3 cell lysates; Mouse colon and P0 bone tissue lysates IHC-P: Human, mouse and rat colon tissue. ICC/IF: SW480 cells and primary hippocampal mouse neurons/glia DIV14. Flow Cyt (intra): PC3 cells. IP: SW480 cell lysate.
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General notes
ab220812 is the carrier-free version of ab185230.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR14335 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab220812 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 56 kDa).
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 70 kDa (predicted molecular weight: 56 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
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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
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Database links
- Entrez Gene: 403464 Dog
- Entrez Gene: 6662 Human
- Entrez Gene: 20682 Mouse
- Entrez Gene: 396840 Pig
- Entrez Gene: 140586 Rat
- Omim: 608160 Human
- SwissProt: Q7YRJ7 Dog
- SwissProt: P48436 Human
see all -
Alternative names
- campomelic dysplasia autosomal sex reversal antibody
- CMD 1 antibody
- CMD1 antibody
see all
Images
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All lanes : Anti-SOX9 antibody [EPR14335] (ab185230) at 1/1000 dilution
Lane 1 : SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 4 : Mouse colon tissue lysate
Lane 5 : Mouse P0 bone tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 56 kDa
Observed band size: 42, 56, 75 kDa why is the actual band size different from the predicted?
Exposure time: 40 secondsThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
Blocking/Diluting buffer and concentration: 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).
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Immunofluorescence staining of SOX9 using ab185230 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 ab185230 at 5 μg/ml, ab4674 (anti-GFAP) at 1/1000 dilution and ab104224 (anti-NeuN) at 1/1000 dilution. Cells were then incubated with ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green), ab150176, Goat Anti-Chicken IgY H&L (Alexa Fluor® 594) preadsorbed (shown in red) and 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.
Images were acquired with the Perkin Elmer Operetta HCA and a maximum intensity projection of confocal sections is shown.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230). -
Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling SOX9 with ab185230 at 1/2000 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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ab185230 at 1/60 immunoprecipitating SOX9 in SW480 (human colorectal adenocarcinoma) whole cell lysate observed at 56 and 75 KDa (lanes 1 and 2).
The expression pattern observed is consistent with the literature (PMID: 10682837) with an additional off-target band at 46 kD.
Lane 1 (input): SW480 whole cell lysate 10μg
Lane 2 (+): SW480 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab185230 in SW480 whole cell lysate
For western blotting, ab185230 (1/1000) was used as the primary antibody and ab131366 VeriBlot for IP (HRP) was used for detection at 1/1000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
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Intracellular flow cytometric analysis of PC3 cells (2% paraformaldehyde-fixed) labeling SOX9 with ab185230 at 1/190 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
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Immunofluorescent analysis of SW480 cells (4% paraformaldehyd-fixed) labeling SOX9 with ab185230 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
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Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling SOX9 with ab185230 at 1/2000 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling SOX9 with ab185230 at 1/2000 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185230).
Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (2)
ab220812 has been referenced in 2 publications.
- Arslan E et al. Glycosaminoglycan-mimetic signals direct the osteo/chondrogenic differentiation of mesenchymal stem cells in a three-dimensional peptide nanofiber extracellular matrix mimetic environment. Biomacromolecules N/A:N/A (2016). Flow Cyt ; Rat . PubMed: 26840042
- Wu Y et al. TrAmplification of Human Dental Follicle Cells by piggyBac Transposon - Mediated Reversible Immortalization System. PLoS One 10:e0130937 (2015). WB ; Human . PubMed: 26172849