Overview

  • Product name

    Human CD73 ELISA Kit (NT5E)
    See all CD73 kits
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    1 16 132pg/ml 5.94 4.5%
    2 16 510pg/ml 28.56 5.6%
    3 16 1427pg/ml 91.33 6.4%
    Inter-assay
    Sample n Mean SD CV%
    1 24 167pg/ml 8.85 5.3%
    2 24 624pg/ml 37.44 6%
    3 24 1553pg/ml 111.82 7.2%
  • Sample type

    Cell culture supernatant, Serum, Cell Lysate, Hep Plasma, EDTA Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    < 10 pg/ml
  • Range

    156 pg/ml - 10000 pg/ml
  • Assay time

    3h 30m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    The Human CD73 Enzyme-Linked Immunosorbent Assay (ELISA) kit (NT5E) (ab213761) is designed for the quantitative measurement of Human CD73 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).


    The ELISA kit is based on standard sandwich enzyme-linked immunosorbent assay technology. A polyclonal antibody from sheep specific for CD73/NT5E has been pre-coated onto 96-well plates. Standards (Expression system for standard: CHO; Immunogen sequence: W27-K547) and test samples are added to the wells, a biotinylated detection polyclonal antibody from sheep specific for CD73/NT5E is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with PBS or TBS buffer. HRP substrate TMB is used to visualize HRP enzymatic reaction. TMB is catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the Human CD73/NT5E amount of sample captured in plate.

  • Notes

    5'-nucleotidase (5'-NT), also known as ecto-5'-nucleotidase or CD73 (Cluster of Differentiation 73), is an enzyme that in humans is encoded by the NT5E gene. Ecto-5-prime-nucleotidase catalyzes the conversion at neutral pH of purine 5-prime mononucleotides to nucleosides, the preferred substrate being AMP. The enzyme consists of a dimer of 2 identical 70-kD subunits bound by a glycosyl phosphatidyl inositol linkage to the external face of the plasma membrane. The enzyme is used as a marker of lymphocyte differentiation. Consequently, a deficiency of NT5 occurs in a variety of immunodeficiency diseases. Other forms of 5-prime nucleotidase exist in the cytoplasm and lysosomes and can be distinguished from ecto-NT5 by their substrate affinities, requirement for divalent magnesium ion, activation by ATP, and inhibition by inorganic phosphate. Rare allelic variants are associated with a syndrome of adult-onset calcification of joints and arteries (CALJA) affecting the iliac, femoral, and tibial arteries reducing circulation in the legs and the joints of the hands and feet causing pain.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at -20°C. Please refer to protocols.
  • Components Identifier 1 x 96 tests
    ABC Diluent Buffer Blue Cap 1 x 12ml
    Adhesive Plate Seal 4 units
    Antibody Diluent Buffer Green Cap 1 x 12ml
    Anti-Human CD73 Coated Microplate (12 x 8 wells) 1 unit
    Avidin-Biotin-Peroxidase Complex (ABC) 1 x 130µl
    Biotinylated anti- Human CD73 Antibody 1 x 130µl
    Lyophilized Recombinant Human CD73 Standard 2 vials
    Sample Diluent Buffer Green Cap 1 x 30ml
    TMB Color Developing Agent Black Cap 1 x 10ml
    TMB Stop Solution Yellow Cap 1 x 10ml
  • Research areas

  • 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
  • Alternative names

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

Applications

Our Abpromise guarantee covers the use of ab213761 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
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Human CD73 ELISA Kit (NT5E) (ab213761) Standard Curve.

Protocols

References

ab213761 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Answer

Plasma samples where citrate has been used as anticoagulant should not be used.

The citrate would interfere with the ELISA.

For each ELISA kit, only the anti-coagulant stated in the product data sheets or protocol booklet should be used.

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