Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Syntenin antibody [EPR8102] (ab133267)

Knockout Tested Rabbit recombinant monoclonal Syntenin antibody [EPR8102]. Validated in WB, IP, IHC, Flow Cyt, ICC/IF and tested in Human. Cited in 9 publication(s). Independently reviewed in 1 review(s).

Overview

  • Product name

    Anti-Syntenin antibody [EPR8102]
    See all Syntenin primary antibodies
  • Description

    Rabbit monoclonal [EPR8102] to Syntenin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, IHC-P, Flow Cyt, 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
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
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.

See IHC antigen retrieval protocols.

Flow Cyt 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.

ICC/IF 1/50 - 1/200.

Target

  • 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
    modifications

    Phosphorylated 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
  • Database links

  • Alternative names

    • MDA-9 antibody
    • MDA9 antibody
    • Melanoma differentiation-associated protein 9 antibody
    • Pro-TGF-alpha cytoplasmic domain-interacting protein 18 antibody
    • Scaffold protein Pbp1 antibody
    • SDCB1_HUMAN antibody
    • SDCBP antibody
    • ST1 antibody
    • SYCL antibody
    • Syndecan binding protein (syntenin) antibody
    • Syndecan binding protein 1 antibody
    • Syndecan-binding protein 1 antibody
    • Syntenin 1 antibody
    • Syntenin-1 antibody
    • TACIP18 antibody
    see all

Images

  • 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.

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Syntenin with purified ab133267 at 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. 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.

  • 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

  • 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 kDa



    Blocking and dilution buffer: 5% NFDM/TBST

  • Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling Syntenin with purified ab133267 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.

  • 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.

     

     

  • 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.

  • 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 kDa



    Blocking and dilution buffer: 5% NFDM/TBST

  • 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 kDa



    Blocking and dilution buffer: 5% NFDM/TBST

  • 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.

  • 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.

  • 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.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:

  • Kordelas L  et al. Individual Immune-Modulatory Capabilities of MSC-Derived Extracellular Vesicle (EV) Preparations and Recipient-Dependent Responsiveness. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30987036) »
  • Liu S  et al. Highly Purified Human Extracellular Vesicles Produced by Stem Cells Alleviate Aging Cellular Phenotypes of Senescent Human Cells. Stem Cells 37:779-790 (2019). Read more (PubMed: 30811771) »
See all 14 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular Smooth Muscle Cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Vascular Smooth Muscle Cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

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Verified customer

Submitted May 30 2018

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