Mouse CD73/NT5E 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 1 203 291 294 288 269 5.92 2.2% Sample 2 987 1055 1152 756 988 19.75 2.0% Sample 3 3240 3451 4010 4526 3807 95.17 2.5% 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 1 203 291 294 288 269 5.92 2.2% Sample 1 203 291 294 288 269 5.92 2.2% Sample 2 987 1055 1152 756 988 19.75 2.0% Sample 2 987 1055 1152 756 988 19.75 2.0% Sample 3 3240 3451 4010 4526 3807 95.17 2.5% Sample 3 3240 3451 4010 4526 3807 95.17 2.5%

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 203 987 3240 274 1150 3163 Standard deviation 8.93 40.47 165.24 15.62 73.60 211.92 CV (%) 4.4% 4.1% 5.1% 5.7% 6.4% 6.7% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 203 987 3240 274 1150 3163 Standard deviation 8.93 40.47 165.24 15.62 73.60 211.92 CV (%) 4.4% 4.1% 5.1% 5.7% 6.4% 6.7% 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) 203 987 3240 274 1150 3163 Mean (pg/ml) 203 987 3240 274 1150 3163 Standard deviation 8.93 40.47 165.24 15.62 73.60 211.92 Standard deviation 8.93 40.47 165.24 15.62 73.60 211.92 CV (%) 4.4% 4.1% 5.1% 5.7% 6.4% 6.7% CV (%) 4.4% 4.1% 5.1% 5.7% 6.4% 6.7%

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. 0.066 0.135 0.218 0.409 0.748 1.281 1.918 2.343 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.066 0.135 0.218 0.409 0.748 1.281 1.918 2.343 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.066 0.135 0.218 0.409 0.748 1.281 1.918 2.343 O.D. 0.066 0.135 0.218 0.409 0.748 1.281 1.918 2.343

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.