Rat Amyloid Beta Peptide 1-42 ELISA Kit

Supplied by Antibodies.com, this competitive enzyme-linked immunosorbent assay (cELISA) is designed for the in vitro quantitative determination of rat Amyloid beta Peptide 1-42 in serum, plasma, or other biological fluids. The 96-well microtiter plate is pre-coated with Amyloid beta Peptide 1-42 antigen. During the incubation, Amyloid beta Peptide 1-42 present in samples or standards competes with the fixed amount of immobilised antigen for binding sites on a biotinylated anti-Amyloid beta Peptide 1-42 antibody; the more analyte present, the less biotinylated antibody that binds to the plate. After washing, an HRP-avidin conjugate is added, and TMB substrate is used to visualise the enzymatic reaction, producing a blue colour that turns yellow upon addition of acidic stop solution. The density of yellow is inversely proportional to Amyloid beta Peptide 1-42 concentration and is read at 450 nm on a colorimetric microplate reader.

The assay is reported as competitive and quantitative, with a stated range of 15.6–1000 pg/ml and a sensitivity of <6.1 pg/ml. Total assay time is 2 h 30 min. Spike-and-recovery performance is reported in serum (average 91%, range 83–100%, n=5), EDTA plasma (average 95%, range 91–99%, n=5) and heparin plasma (average 94%, range 90–99%, n=5). Dilution linearity across 1:2 to 1:16 dilutions in the same three matrices is also reported.

Kit components include a pre-coated 96-well microplate (12 × 8 well strips), lyophilised standard (2 vials), Detection Solution A (70 µl), Detection Solution B (120 µl), 30× Wash Buffer (20 ml), Sample Dilution Buffer (45 ml), TMB Substrate (9 ml), Stop Solution (6 ml) and adhesive plate sealers. Standards, detection solutions and sample dilution buffer are stored at -20 °C, while wash buffer, TMB substrate and stop solution are stored at +4 °C. Components should not be used past the kit's stated expiration date.