Overview

  • Product name

    Anti-CD73 antibody [EPR6114]
    See all CD73 primary antibodies
  • Description

    Rabbit monoclonal [EPR6114] to CD73
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Synthetic peptide, corresponding to an internal sequence within amino acids 50-100 of Human CD73 (UniProt P21589).

  • Positive control

    • WB: Human fetal liver tissue lysate; A375 and HepG2 whole cell lysate (ab7900) IHC-P: Human lung carcinoma, Human tonsil tissue ICC/IF: A375 cells
  • 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 ab133582 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
ICC/IF 1/250.

For unpurified use ad 1/50 - 1/100

IHC-P 1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

For unpurified use ad 1/100 - 1/250. See IHC antigen retrieval protocols.

WB 1/1000 - 1/10000. Predicted molecular weight: 63 kDa.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function

      Hydrolyzes extracellular nucleotides into membrane permeable nucleosides.
    • Involvement in disease

      Defects in NT5E are the cause of calcification of joints and arteries (CAJA) [MIM:211800]. A condition characterized by adult-onset calcification of the lower extremity arteries, including the iliac, femoral and tibial arteries, and hand and foot capsule joints. Age of onset has been reported as early as the second decade of life, usually involving intense joint pain or calcification in the hands.
    • Sequence similarities

      Belongs to the 5'-nucleotidase family.
    • Cellular localization

      Cell membrane.
    • Information by UniProt
    • Database links

    • Alternative names

      • 5' NT antibody
      • 5' nucleotidase (CD73) antibody
      • 5' nucleotidase precursor antibody
      • 5' nucleotidase, ecto antibody
      • 5' nucleotidase, ecto (CD73) antibody
      • 5'-NT antibody
      • 5'-nucleotidase antibody
      • 5NTD_HUMAN antibody
      • CD73 antibody
      • CD73 antigen antibody
      • E5NT antibody
      • Ecto 5' nucleotidase antibody
      • Ecto-5'-nucleotidase antibody
      • eN antibody
      • eNT antibody
      • NT antibody
      • NT5 antibody
      • NT5E antibody
      • NTE antibody
      • Purine 5 Prime Nucleotidase antibody
      see all

    Images

    • All lanes : Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution

      Lane 1 : Wild-type HEK-293 whole cell lysate
      Lane 2 : NT5E knockout HEK-293 whole cell lysate
      Lane 3 : A375 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 63 kDa



      Lanes 1 - 3: Merged signal (red and green). Green - ab133582 observed at 63 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab133582 was shown to recognize NT5E in wild-type HEK-293 cells as signal was lost at the expected MW in NT5E knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NT5E knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab133582 and ab8245 (Mouse anti-GAPDH 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung carcinoma tissue sections labeling CD73 with Purified ab133582 at 1:2000 dilution (1.12 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used. Negative control:PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
    • Immunocytochemistry/ Immunofluorescence analysis of A375 (Human malignant melanoma epithelial cell) cells labeling CD73 with Purified ab133582 at 1:250 dilution (8.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
    • Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution + A375 ( Human malignant melanoma epithelial cell) whole cell lysates at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 63 kDa

    • Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling CD73 with ab133582 at 1/100 dilution.
    • All lanes : Anti-CD73 antibody [EPR6114] (ab133582) at 1/1000 dilution

      Lane 1 : Human fetal liver tissue lysate
      Lane 2 : A375 cell lysate
      Lane 3 : HepG2 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution

      Predicted band size: 63 kDa

    References

    This product has been referenced in:

    • Ikuta D  et al. Fibrosis in metastatic lymph nodes is clinically correlated to poor prognosis in colorectal cancer. Oncotarget 9:29574-29586 (2018). Read more (PubMed: 30038705) »
    • Kawasaki T  et al. Six1 is required for mouse dental follicle cell and human periodontal ligament-derived cell proliferation. Dev Growth Differ N/A:N/A (2016). IF . Read more (PubMed: 27241908) »
    See all 5 Publications for this product

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (T84)
    Gel Running Conditions
    Non-reduced Denaturing
    Loading amount
    1e+006 cells
    Specification
    T84
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Oct 19 2018

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