Human Forkhead Box Protein O1 ELISA Kit

A sandwich enzyme-linked immunosorbent assay (sELISA) supplied by Antibodies.com, this kit is designed for the in vitro quantitative determination of human FOXO1A in tissue homogenates, cell lysates, and other biological fluids. The assay uses the sandwich enzyme immunoassay technique, in which an antibody specific for FOXO1A is pre-coated onto a microplate. Standards and samples are added to the wells and bound by the immobilised antibody, followed by incubation with a biotinylated anti-FOXO1A antibody and then an HRP-avidin conjugate. A TMB substrate solution is used to visualise the enzymatic reaction, producing a blue product that turns yellow upon addition of the stop solution; absorbance is read at 450 nm in a microplate reader, and FOXO1A concentration is calculated by reference to the standard curve.

The assay has a reported sensitivity of 0.046 ng/ml and a quantitative range of 0.156–10 ng/ml, with a total assay time of 4 hours 30 minutes. Detection is colorimetric and intended for use with a microplate reader. In spiked recovery testing (n=5 per matrix), serum samples showed recoveries of 80%–102% (average 91%), EDTA plasma 81%–100% (average 90%), and heparin plasma 80%–89% (average 84%). Dilution linearity has been demonstrated at 1:2, 1:4, 1:8, and 1:16 dilutions, with recoveries of 74%–107% for serum, 79%–108% for EDTA plasma, and 82%–105% for heparin plasma. Kit components should be stored at +4°C or –20°C as instructed and not used past the expiration date.