Human Nicotinamide Adenine Dinucleotide Phosphate Oxidase 4 ELISA Kit

A sandwich Enzyme-Linked Immunosorbent Assay (sELISA) from Antibodies.com, designed for the in vitro quantitative determination of human NADPH Oxidase 4 in tissue homogenates, cell lysates, or other biological fluids. Reactivity is human and the detection is colorimetric, read on a microplate reader at 450 nm.

The assay employs a pre-coated 96-well microplate (12 x 8 well strips) immobilised with an antibody specific for NADPH Oxidase 4. Standards and test samples are added to the wells, followed by a Biotinylated Anti-NADPH Oxidase 4 Antibody and an HRP-Avidin conjugate. A TMB substrate solution is used to visualise the HRP enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution; the yellow intensity is proportional to the NADPH Oxidase 4 captured, and concentration is calculated from a standard curve. Total assay time is 4 hours 30 minutes. The stated sensitivity is <0.051 ng/ml and the assay range is 0.156–10 ng/ml.

Kit components should be stored at +4°C or -20°C as instructed, and the kit must not be used past its expiration date. Reported recovery ranges across dilutions of 1:2, 1:4, 1:8 and 1:16 are: Serum (n=5) 87–91%, 87–107%, 74–101%, 92–97%; EDTA Plasma (n=5) 90–105%, 84–101%, 90–101%, 79–108%; Heparin Plasma (n=5) 84–95%, 92–105%, 82–105%, 89–91%. Average recoveries are: Serum 80–102% (91%), EDTA Plasma 81–100% (90%), Heparin Plasma 80–89% (84%).