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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

  • Datasheet
  • Certificate of Compliance
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Western blot - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Immunoprecipitation - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Flow Cytometry - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Flow Cytometry - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • OI-RD Scanning - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
  • Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

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 - BSA and Azide free
  • Suitable for: IHC-P, IP, ICC/IF, Flow Cyt, WB
  • Knockout validated
  • Reacts with: Human

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Conjugation
Product image
HRP Conjugation Kit - Lightning-Link® (ab102890)
Protein
Recombinant Human Syntenin protein (ab185832)

View more associated products

Overview

  • Product name

    Anti-Syntenin antibody [EPR8102] - BSA and Azide free
    See all Syntenin primary antibodies
  • Description

    Rabbit monoclonal [EPR8102] to Syntenin - BSA and Azide free
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, A549 and HeLa cell lysates.
  • General notes

    ab236071 is the carrier-free version of ab133267 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    Ab236071 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    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
    For more information see here.

    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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Dissociation constant (KD)

    KD = 1.24 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.2
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8102
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Synapse marker
    • Neuroscience
    • Neurology process
    • Neurogenesis

Associated products

  • Alternative Versions

    • Anti-Syntenin antibody [EPR8102] (ab133267)
    • Alexa Fluor® 647 Anti-Syntenin antibody [EPR8102] (ab205772)
    • PE Anti-Syntenin antibody [EPR8102] (ab210837)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • HeLa whole cell lysate (ab29545)
    • HeLa membrane extract lysate (ab29547)
    • A549 whole cell lysate (ab7910)
  • Recombinant Protein

    • Recombinant Human Syntenin protein (ab185832)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab236071 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Human
Application Abreviews Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab172730 - 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 32 kDa (predicted molecular weight: 32 kDa).
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

ab172730 - 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 32 kDa (predicted molecular weight: 32 kDa).

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.
  • Target information above from: UniProt accession O00560 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6386 Human
    • Omim: 602217 Human
    • SwissProt: O00560 Human
    • Unigene: 200804 Human
    • 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

    • Western blot - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Western blot - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Immunoprecipitation - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Immunoprecipitation - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

       

       

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Flow Cytometry - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Flow Cytometry - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Immunocytochemistry/ Immunofluorescence - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Flow Cytometry - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Flow Cytometry - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

      Immunohistochemical analysis of Syntenin in paraffin embedded Human brain tissue, using unpurified ab133267 at a 1/50 dilution.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • OI-RD Scanning - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      OI-RD Scanning - Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133267).

    • Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)
      Anti-Syntenin antibody [EPR8102] - BSA and Azide free (ab236071)

    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.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

  • References (0)

    Publishing research using ab236071? Please let us know so that we can cite the reference in this datasheet.

    ab236071 has not yet been referenced specifically in any publications.

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