Recombinant Anti-Syntenin antibody [EPR8102] (ab133267)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8102] to Syntenin
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Syntenin antibody [EPR8102]
See all Syntenin primary antibodies -
Description
Rabbit monoclonal [EPR8102] to Syntenin -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Syntenin aa 1-100. The exact sequence is proprietary.
Database link: O00560 -
Positive control
- Human fetal brain lysate, Human fetal heart lysate, Human placenta lysate, HeLa, 293T and A549 (Human lung carcinoma cell line) cell lysates; Human brain tissue
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 1.24 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR8102 -
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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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133267 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) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 - 1/10000. |
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WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).
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IP |
1/10 - 1/100.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
1/50 - 1/200.
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Notes |
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Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 - 1/10000. |
WB
1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/50 - 1/200. |
Target
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Function
Seems to function as an adapter protein. In adherens junctions may function to couple syndecans to cytoskeletal proteins or signaling components. Seems to couple transcription factor SOX4 to the IL-5 receptor (IL5RA). May also play a role in vesicular trafficking. Seems to be required for the targeting of TGFA to the cell surface in the early secretory pathway. -
Tissue specificity
Widely expressed. Expressed in fetal kidney, liver, lung and brain. In adult highest expression in heart and placenta. -
Sequence similarities
Contains 2 PDZ (DHR) domains. -
Post-translational
modificationsPhosphorylated on tyrosine residues. -
Cellular localization
Cell junction > focal adhesion. Cell junction > adherens junction. Cell membrane. Endoplasmic reticulum membrane. Nucleus. Melanosome. Cytoplasm > cytosol. Cytoplasm > cytoskeleton. Mainly membrane-associated. Localized to adherens junctions, focal adhesions and endoplasmic reticulum. Colocalized with actin stress fibers. Also found in the nucleus. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 6386 Human
- Omim: 602217 Human
- SwissProt: O00560 Human
- Unigene: 200804 Human
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Alternative names
- MDA-9 antibody
- MDA9 antibody
- Melanoma differentiation-associated protein 9 antibody
see all
Images
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Syntenin knockout HAP1 whole cell lysate (20 µg)
Lane 3: A549 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab133267 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab133267 was shown to recognize Syntenin in wild-type HAP1 cells as signal was lost at the expected MW in Syntenin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Syntenin knockout samples were subjected to SDS-PAGE. ab133267 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular carcinoma tissue labeling Syntenin with purified ab133267 at 1/50 dilution. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin
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Anti-Syntenin antibody [EPR8102] (ab133267) at 1/2000 dilution (purified) + A549 (Human lung carcinoma cell line) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and dilution buffer: 5% NFDM/TBST
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Intracellular Flow Cytometry analysis ofHeLa (Human epithelial cell line from cervix adenocarcinoma)labeling Syntenin with purifiedab133267 at 1/50 (red).Cells were fixed with 4% paraformaldehyde. A goat anti rabbit IgG (Alexa Fluor® 488) 1/2000 was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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ab133267 (purified) at 1/40 immunoprecipitating Syntenin in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab133267 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133267 in HeLa whole cell lysate.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebral cortex tissue labeling Syntenin with purified ab133267 at 1/50 dilution. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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All lanes : Anti-Syntenin antibody [EPR8102] (ab133267) at 1/2000 dilution (purified)
Lane 1 : Human placenta lysate
Lane 2 : Human fetal brain lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and dilution buffer: 5% NFDM/TBST
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Anti-Syntenin antibody [EPR8102] (ab133267) at 1/10000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and dilution buffer: 5% NFDM/TBST
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ab133267 staining Syntenin in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.
Negative control 1: PBS only.
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Overlay histogram showing SHSY-5Y cells stained with unpurified ab133267 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133267, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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All lanes : Anti-Syntenin antibody [EPR8102] (ab133267) at 1/1000 dilution (unpurified)
Lane 1 : Human fetal brain lysate
Lane 2 : 293T cell lysate
Lane 3 : Human fetal heart lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa -
Immunohistochemical analysis of Syntenin in paraffin embedded Human brain tissue, using unpurified ab133267 at a 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (68)
ab133267 has been referenced in 68 publications.
- Choezom D & Gross JC Neutral sphingomyelinase 2 controls exosome secretion by counteracting V-ATPase-mediated endosome acidification. J Cell Sci 135:N/A (2022). PubMed: 35050379
- Scott TA et al. Engineered extracellular vesicles directed to the spike protein inhibit SARS-CoV-2. Mol Ther Methods Clin Dev 24:355-366 (2022). PubMed: 35127966
- Li DK et al. Exosomal HMGA2 protein from EBV-positive NPC cells destroys vascular endothelial barriers and induces endothelial-to-mesenchymal transition to promote metastasis. Cancer Gene Ther 29:1439-1451 (2022). PubMed: 35388172
- Chetty VK et al. Efficient Small Extracellular Vesicles (EV) Isolation Method and Evaluation of EV-Associated DNA Role in Cell-Cell Communication in Cancer. Cancers (Basel) 14:N/A (2022). PubMed: 35565197
- Shi Y et al. Huaier Inhibits Gastric Cancer Growth and Hepatic Metastasis by Reducing Syntenin Expression and STAT3 Phosphorylation. J Oncol 2022:6065516 (2022). PubMed: 35756080