Recombinant Anti-SOX9 antibody [EPR14335-78] - BSA and Azide free (ab225541)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR14335-78] to SOX9 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SOX9 antibody [EPR14335-78] - BSA and Azide free
See all SOX9 primary antibodies -
Description
Rabbit monoclonal [EPR14335-78] to SOX9 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore 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
- 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, primary hippocampal mouse neurons/glia DIV14. Flow Cyt (intra): PC-3 cells.
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General notes
ab225541 is the carrier-free version of ab185966.
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.
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
EPR14335-78 -
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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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab225541 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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ICC/IF |
Use at an assay dependent concentration.
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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 citrate buffer. |
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. |
ICC/IF
Use at an assay dependent concentration. |
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 citrate buffer. |
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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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, 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. 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185966).
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Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded pig small intestine sections labelling 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185966).
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All lanes : Anti-SOX9 antibody [EPR14335-78] (ab185966) 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: 180 secondsThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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).
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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 permeabilised 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185966).
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Immunohistochemistry analysis of paraffin-embedded Human breast carcinoma tissue labeling SOX9 with ab185966 at 1/1000 dilution. Counterstained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185966).
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Immunofluorescence analysis of 4% paraformaldehyde fixed SW480 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185966).
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Immunofluorescence analysis of 4% paraformaldehyde fixed PC3 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185966).
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This ICC data was generated using the same anti-SOX9 antibody clone [EPR14335-78] in a different buffer formulation (cat# ab185966).
ab185966 staining Sox9 in F9 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 5μg/ml concentration and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin, at 1μg/ml concentration, followed by a further incubation at room temperature for 1h 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 labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab225541 has been referenced in 1 publication.
- Shi J et al. Deubiquitinase USP47/UBP64E Regulates ß-Catenin Ubiquitination and Degradation and Plays a Positive Role in Wnt Signaling. Mol Cell Biol 35:3301-11 (2015). PubMed: 26169834