Human NQO1 ELISA Kit

Manufactured by Assay Genie, this sandwich ELISA kit is designed for the quantitative measurement of NQO1 (NAD(P)H dehydrogenase quinone 1; UniProt P15559) in human samples using a colorimetric, double-antibody detection format. The assay is validated for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples, and is intended for cell biology research where NQO1 levels are studied in relation to quinone detoxification and protection against oxidative stress.

Sensitivity is reported as 0.188 ng/mL with a measurable range of 0.313–20 ng/mL. The assay follows a six-step workflow: reagent and plate equilibration to room temperature; primary incubation at 37°C for 90 minutes; biotinylated detection antibody binding at 37°C for 60 minutes; HRP-streptavidin binding at 37°C for 30 minutes; TMB colour development in the dark for 10–20 minutes; and absorbance reading at 450 nm immediately after stop solution is added. Sample preparation guidance covers serum (allow to clot, centrifuge at 1000 × g for 20 minutes), plasma (collect in anticoagulant tubes, centrifuge at 1000 × g for 15 minutes at 2–8°C), tissue homogenate in PBS with protease inhibitors, cell culture supernatant (centrifuge at 2500 rpm for 5 minutes), and cell lysates.

Reported linearity at 1:2, 1:4 and 1:8 dilutions is 85–105%, 92–96% and 90–105% for serum (n = 5); 88–99%, 83–100% and 83–98% for EDTA plasma (n = 5); and 87–100%, 82–100% and 88–93% for heparin plasma (n = 5). Recovery ranges are 85–100% with an average of 92% for serum, 86–94% with an average of 92% for EDTA plasma, and 88–104% with an average of 98% for heparin plasma. The kit should be stored at 2–8°C, at which temperature the unopened components remain stable for 12 months.