Human Neutrophil Gelatinase Associated Lipocalin ELISA Kit

From Antibodies.com, this sandwich ELISA kit is designed for the in vitro quantitative determination of human Lipocalin-2/NGAL (Neutrophil Gelatinase Associated Lipocalin) in serum, plasma, urine, tissue homogenates, cell lysates, cell culture supernates or other biological fluids. The assay employs the sandwich enzyme immunoassay technique: an antibody specific for Lipocalin-2/NGAL is pre-coated onto a 96-well microplate (12 x 8 well strips), and bound analyte is detected using a Biotinylated Anti-Lipocalin-2/NGAL Antibody followed by an HRP-Avidin conjugate. TMB substrate is added and, after incubation with the acidic stop solution, the resulting yellow product is read at 450 nm, with colour intensity proportional to the amount of Lipocalin-2/NGAL captured in each well.

Quantitative range is 0.156–10 ng/ml with a sensitivity of <0.06 ng/ml, and the protocol takes approximately 4 h 30 min. Spiked recovery averages 91% in serum (range 80–102%), 90% in EDTA plasma (range 81–100%) and 84% in heparin plasma (range 80–89%). The kit contains a pre-coated 96-well microplate, two vials of lyophilised standard, Detection Solution A (120 µl), Detection Solution B (120 µl), 30X Wash Buffer (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml) and plate sealers (5 adhesive strips); components are stored at +4 °C or –20 °C as indicated, and the kit is intended for use before its expiry date with readings taken on a colorimetric microplate reader.