Human Nephronectin ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human Nephronectin in tissue homogenates or other biological fluids. The microplate is pre-coated with an antibody specific for Nephronectin. Standards and test samples are added to the wells so that the analyte is captured by the immobilised antibody. After washing, a Biotinylated Anti-Nephronectin Antibody binds the captured target, followed by incubation with an HRP-Avidin conjugate. A further wash precedes addition of TMB substrate; the resulting HRP-catalysed reaction produces a blue-coloured product that turns yellow on addition of an acidic stop solution. Absorbance is read at 450 nm using a colorimetric microplate reader, and the Nephronectin concentration is calculated by reference to a standard curve.

Key specifications include a quantitative assay range of 0.156–10 ng/ml and a sensitivity of <0.054 ng/ml, with a total assay time of 4h 30m. Spike-and-recovery performance has been validated in serum, EDTA plasma, and heparin plasma (n=5 each), giving recoveries of 80%–102% (average 91%) in serum, 81%–100% (average 90%) in EDTA plasma, and 80%–89% (average 84%) in heparin plasma. Dilution linearity has been assessed at 1:2, 1:4, 1:8, and 1:16 dilutions in each matrix (n=5). Detection is colourimetric, and kit components should be stored at +4°C or −20°C as instructed.