Human Nitric Oxide Synthase 2, Inducible ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human inducible nitric oxide synthase (iNOS, also known as Nitric Oxide Synthase 2, Inducible) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.

An antibody specific for iNOS has been pre-coated onto a 96-well microtiter plate. Standards and test samples are added and the iNOS present binds to the immobilised antibody. Following incubation and washing, a Biotinylated Anti-iNOS Antibody binds the captured iNOS, and HRP-Avidin conjugate is then added. After further incubation and washing, TMB substrate solution is used to visualise the HRP enzymatic reaction, producing a blue product that changes to yellow upon addition of acidic stop solution. The intensity of yellow is proportional to the amount of iNOS captured per well, and the concentration is calculated by reading the absorbance at 450 nm in a microplate reader against a standard curve.

The quantitative assay has a product range of 0.156–10 ng/ml and a sensitivity of <0.064 ng/ml, with an assay time of 4 hours 30 minutes on a colorimetric microplate reader platform. Components supplied include a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard (2 vials), detection solutions A and B (120 µl each), 30X wash buffer (20 ml), sample dilution buffer (45 ml), TMB substrate (9 ml), stop solution (6 ml) and plate sealers (5 adhesive strips). Reported sample recovery averages are 91% for serum, 90% for EDTA plasma and 84% for heparin plasma (each n=5), and linearity has been demonstrated at sample dilutions of 1:2, 1:4, 1:8 and 1:16.