Human CD73 ELISA Kit (NT5E)

製品の詳細

Human CD73 ELISA Kit (NT5E) ab213761 is a sandwich ELISA to measure Human CD73 (NT5E) in serum, EDTA plasma, heparin plasma, cell culture supernatant, cell lysate with a sensitivity of 10 pg/ml.

How the assay works

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.

Assay Specificity

Our ELISA kits are rigorously validated to ensure the highest level of consistency and reproducibility. Please check the protocol booklet for more details

Human CD73 ELISA Kit (NT5E) ab213761 protocol summary

1. Add standard or sample to each well used. Incubate 37C
2. Add prepared biotin antibody to each well. Incubate at 37C
3. Add prepared Avidin-BiotinPeroxidase Complex (ABC). Incubate at 37C
4. Add TMB to each well. Incubate at room temperature.
5. Add Stop Solution to each well. Read at 450 nm

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.