Human ISL LIM Homeobox Protein 1 ELISA Kit

Antibodies.com provides this sandwich ELISA (sELISA) intended for the in vitro quantitative determination of human Islet 1 (ISL LIM Homeobox Protein 1) in tissue homogenates or other biological fluids. The assay employs a 96-well microplate pre-coated with an antibody specific for Islet 1. Standards and test samples are added to the wells, where Islet 1 is captured by the immobilised antibody. After washing, a Biotinylated Anti-Islet 1 Antibody binds the captured analyte, and an HRP-Avidin conjugate is added. TMB substrate solution is then used to visualise the enzymatic reaction, producing a blue product that turns yellow upon addition of acidic stop solution. The intensity of yellow is proportional to the amount of Islet 1 captured, and concentration is calculated from O.D. absorbance at 450 nm against a standard curve.

The assay has a stated range of 0.156–10 ng/ml and a sensitivity of <0.063 ng/ml, with a run time of 4h 30m. Detection is colorimetric and is read on a microplate reader. Sample recovery averages are reported as: serum 91% (range 80–102%, n=5), EDTA plasma 90% (range 81–100%, n=5), and heparin plasma 84% (range 80–89%, n=5). Dilution linearity was demonstrated across 1:2, 1:4, 1:8, and 1:16 dilutions in serum, EDTA plasma, and heparin plasma (n=5).

Kit components comprise a pre-coated 96-well microplate (12 x 8 well strips), lyophilised standard (2 vials), 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 5 adhesive plate sealers. Components are stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.