Human CD73/NT5E/Ecto-5'-nucleotidase EZ-Set™ ELISA Kit (DIY Antibody Pairs)

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.

Catalog number Description Quantity Storage of opened/reconstituted material EZ1422-CA Sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1422-DA Biotinylated sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1422-ST Lyophilized recombinant human CD73/NT5E/Ecto-5'-nucleotidase standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material EZ1422-CA Sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1422-DA Biotinylated sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1422-ST Lyophilized recombinant human CD73/NT5E/Ecto-5'-nucleotidase standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. Catalog number Description Quantity Storage of opened/reconstituted material Catalog number Description Quantity Storage of opened/reconstituted material EZ1422-CA Sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1422-CA Sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Capture Antibody) 500 μl, 4 μg/mL (recommended dilution 1:100) Store undiluted at 4°C for 1 month or at -20°C for 3 months provided this is within the expiration date of the kit. EZ1422-DA Biotinylated sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) EZ1422-DA Biotinylated sheep anti-human CD73/NT5E/Ecto-5'-nucleotidase polyclonal antibody (Detection Antibody) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) AR1103 Avidin-Biotin-Peroxidase Complex (ABC) 500 μl (recommended dilution 1:100) EZ1422-ST Lyophilized recombinant human CD73/NT5E/Ecto-5'-nucleotidase standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit. EZ1422-ST Lyophilized recombinant human CD73/NT5E/Ecto-5'-nucleotidase standard 10 ng/tubex3 Discard the standard stock solution after 12 hours at 4°C. May be stored at -20°C for 48 hours provided this is within the expiration date of the kit.

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. 0.055 0.125 0.169 0.299 0.545 0.956 1.437 2.064 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.055 0.125 0.169 0.299 0.545 0.956 1.437 2.064 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 Concentration (pg/ml) 0 156 312 625 1250 2500 5000 10000 O.D. 0.055 0.125 0.169 0.299 0.545 0.956 1.437 2.064 O.D. 0.055 0.125 0.169 0.299 0.545 0.956 1.437 2.064

1. Microplate reader in standard size. 2. Automated plate washer. 3. Incubator. 4. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. 5. Clean tubes and Eppendorf tubes. 6. 96 well microplate (Cat# AR1100) 7. Plate Sealers. 8. Capture Antibody Diluent: PBS. 9. Reagent Diluent: 1% BSA in PBS, pH 7.2-7.4, 0.2 μm filtered. 10. Substrate Solution: Tetramethylbenzidine (Cat# AR1104) 11. Stop Solution: 2 N H2SO4 (Cat# AR1105) 12. Wash Buffer (PBS and PBS-T) . Recipes: PBS: 8g NaCl, 0.2g KCl, 1.15g Na2HPO4, 0.2g KH2PO4, adjust the total volume to 1 L with distilled water, pH 7.2-7.4, 0.2 μm filtered. PBS-T: 0.1% Tween® 20 in PBS, pH 7.2-7.4. *Item 6 – 12 are included in the EZ Set Accessory Kit (EZA001)