Human CD73/NT5E/Ecto-5'-nucleotidase ELISA Kit PicoKine®

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. And 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. Rareallelic 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.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 230 235 243 260 242 11.37 4.6% Sample 2 1941 1870 1830 1891 1883 40.01 2.1% Sample 3 5361 4741 5455 5425 5245 293.24 5.5% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 230 235 243 260 242 11.37 4.6% Sample 1 230 235 243 260 242 11.37 4.6% Sample 2 1941 1870 1830 1891 1883 40.01 2.1% Sample 2 1941 1870 1830 1891 1883 40.01 2.1% Sample 3 5361 4741 5455 5425 5245 293.24 5.5% Sample 3 5361 4741 5455 5425 5245 293.24 5.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 230 1941 5361 222 1912 4929 Standard deviation 15.41 79.58 387.29 18.64 101.33 359.81 CV (%) 6.7% 4.1% 7.2% 8.4% 5.3% 7.3% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 230 1941 5361 222 1912 4929 Standard deviation 15.41 79.58 387.29 18.64 101.33 359.81 CV (%) 6.7% 4.1% 7.2% 8.4% 5.3% 7.3% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 230 1941 5361 222 1912 4929 Mean (pg/ml) 230 1941 5361 222 1912 4929 Standard deviation 15.41 79.58 387.29 18.64 101.33 359.81 Standard deviation 15.41 79.58 387.29 18.64 101.33 359.81 CV (%) 6.7% 4.1% 7.2% 8.4% 5.3% 7.3% CV (%) 6.7% 4.1% 7.2% 8.4% 5.3% 7.3%

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.056 0.099 0.132 0.201 0.348 0.6254 1.135 1.946 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.056 0.099 0.132 0.201 0.348 0.6254 1.135 1.946 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.056 0.099 0.132 0.201 0.348 0.6254 1.135 1.946 O.D. 0.056 0.099 0.132 0.201 0.348 0.6254 1.135 1.946

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.