Recombinant Anti-SOX17 antibody [EPR20684] (ab224637)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20684] to SOX17
- Suitable for: ICC/IF, IHC-P, IP, WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-SOX17 antibody [EPR20684]
See all SOX17 primary antibodies -
Description
Rabbit monoclonal [EPR20684] to SOX17 -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, IHC-P, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, SK-OV-3 and OVCAR-3 cell lysates; SOX17 recombinant protein; Rat E4 embryo, E9.5 embryo and rat lung tissue lysates. IHC-P: Human seminoma tissue; Rat lung tissue; Mouse spleen tissue. IP: SK-OV-3 whole cell lysate. ICC/IF: HeLa cells.
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General notes
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 short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20684 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab224637 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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ICC/IF | (2) |
Use a concentration of 0.2 µg/ml.
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IHC-P | (1) |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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WB |
1/500. Detects a band of approximately 55 kDa (predicted molecular weight: 44 kDa).
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Notes |
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ICC/IF
Use a concentration of 0.2 µg/ml. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
WB
1/500. Detects a band of approximately 55 kDa (predicted molecular weight: 44 kDa). |
Target
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Function
Acts as transcription regulator that binds target promoter DNA and bends the DNA. Binds to the sequences 5'-AACAAT-'3 or 5'-AACAAAG-3'. Modulates transcriptional regulation via WNT3A. Inhibits Wnt signaling. Promotes degradation of activated CTNNB1. Plays a key role in the regulation of embryonic development. Required for normal looping of the embryonic heart tube. Required for normal development of the definitive gut endoderm. Probable transcriptional activator in the premeiotic germ cells. -
Tissue specificity
Expressed in adult heart, lung, spleen, testis, ovary, placenta, fetal lung, and kidney. In normal gastrointestinal tract, it is preferentially expressed in esophagus, stomach and small intestine than in colon and rectum. -
Involvement in disease
Defects in SOX17 are the cause of vesicoureteral reflux type 3 (VUR3) [MIM:613674]. VUR3 is a disease belonging to the group of congenital anomalies of the kidney and urinary tract. It is characterized by the reflux of urine from the bladder into the ureters and sometimes into the kidneys, and is a risk factor for urinary tract infections. Primary disease results from a developmental defect of the ureterovesical junction. In combination with intrarenal reflux, the resulting inflammatory reaction may result in renal injury or scarring, also called reflux nephropathy. Extensive renal scarring impairs renal function and may predispose patients to hypertension, proteinuria, renal insufficiency and end-stage renal disease. -
Sequence similarities
Contains 1 HMG box DNA-binding domain.
Contains 1 Sox C-terminal domain. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 64321 Human
- Entrez Gene: 20671 Mouse
- Entrez Gene: 312936 Rat
- Omim: 610928 Human
- SwissProt: Q9H6I2 Human
- SwissProt: Q61473 Mouse
- Unigene: 98367 Human
- Unigene: 279103 Mouse
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Alternative names
- FLJ22252 antibody
- SOX17 antibody
- SOX17_HUMAN antibody
see all
Images
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ab224637 staining SOX17 in wild-type HeLa cells (top panel) and SOX17 knockout HeLa cells (ab265744) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then 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 with ab224637 at 0.2μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] (ab224637)
Immunohistochemical analysis of paraffin-embedded human seminoma tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on tumor cells of human seminoma (PMID:19369635; PMID:18348160) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-SOX17 antibody [EPR20684] (ab224637) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SOX17 knockout HeLa cell lysate
Lane 3 : SK-OV-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 44 kDa
Observed band size: 51 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab224637 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa.
ab224637 Anti-SOX17 antibody [EPR20684] was shown to specifically react with SOX17 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265744 (knockout cell lysate ab257697) was used. Wild-type and SOX17 knockout samples were subjected to SDS-PAGE. ab224637 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-SOX17 antibody [EPR20684] (ab224637) at 1/500 dilution
Lane 1 : SK-OV-3 (Human adenocarcinoma) whole cell lysates with NFDM/TBST
Lane 2 : NIH: OVCAR-3 (Human ovary adenocarcinoma) whole cell lysates with NFDM/TBST
Lane 3 : HeLa (Human cervix adenocarcinoma) whole cell lysates with NFDM/TBST
Lane 4 : Rat E14 embryo tissue lysates with NFDM/TBST
Lane 5 : Rat E9.5 embryo tissue lysates with NFDM/TBST
Lane 6 : Rat lung tissue lysates with NFDM/TBST
Lysates/proteins at 20 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG) at 1/2000 dilution
Predicted band size: 44 kDaExposure time:
Lanes 1-2: 5 seconds
Lanes 3-6: 3 minutesThe expression profile observed is consistent with the literature (PMID: 11786926).
Negative control: HeLa.
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SOX17 was immunoprecipitated from 0.35 mg of SK-OV-3 (human ovarian cancer cell line) lysate with ab224637 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224637 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: SK-OV-3 whole cell lysate 10 µg (Input).
Lane 2: ab224637 IP in SK-OV-3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224637 in SK-OV-3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] (ab224637)
Immunohistochemical analysis of paraffin-embedded human choriocarcinoma tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Negative tissue: no staining on tumor cells of human choriocarcinoma (PMID: 19369635) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] (ab224637)
Immunohistochemical analysis of paraffin-embedded rat lung tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on endothelium of rat lung (PMID:24418654) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOX17 antibody [EPR20684] (ab224637)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling SOX17 with ab224637 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Nuclear staining on endothelium of mouse spleen (PMID:24418654) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab224637 has been referenced in 3 publications.
- Yang Y et al. MicroRNA let-7i Inhibits Histone Lysine Demethylase KDM5B to Halt Esophageal Cancer Progression. Mol Ther Nucleic Acids 22:846-861 (2020). PubMed: 33230480
- Hillel-Karniel C et al. Multi-lineage Lung Regeneration by Stem Cell Transplantation across Major Genetic Barriers. Cell Rep 30:807-819.e4 (2020). PubMed: 31968255
- Chew LJ et al. Sox17 Regulates a Program of Oligodendrocyte Progenitor Cell Expansion and Differentiation during Development and Repair. Cell Rep 29:3173-3186.e7 (2019). PubMed: 31801081