Human Kinesin Family, Member 5A ELISA Kit

Antibodies.com supplies this Sandwich ELISA kit for the in vitro quantitative determination of human KIF5A in tissue homogenates, cell lysates or other biological fluids. The format uses a pre-coated 96-well microplate (12 x 8 well strips) with a colourmetric readout read at 450 nm on a microplate reader. KIF5A bound by the immobilised capture antibody is detected with a Biotinylated Anti-KIF5A Antibody followed by an HRP-Avidin conjugate and TMB substrate; the enzymatic reaction is stopped with an acidic stop solution and the absorbance is compared with a standard curve to calculate sample concentration.

Quantitative performance figures from the manufacturer state an assay range of 0.312–20 ng/ml and a sensitivity of <0.118 ng/ml. Spike-and-recovery in human samples (n=5) was reported as 83–100% (serum, average 91%), 90–99% (EDTA plasma, average 94%) and 91–101% (heparin plasma, average 96%). Dilution linearity across 1:2, 1:4, 1:8 and 1:16 was 90–101% / 80–108% / 84–95% / 92–104% for serum, 82–105% / 89–91% / 87–109% / 74–104% for EDTA plasma and 90–94% / 87–108% / 86–101% / 90–101% for heparin plasma. Total assay time is 4h 30m. Supplied components comprise the pre-coated strip plate, 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 plate sealers (5 adhesive strips), with components stored at +4°C or -20°C as instructed and the kit not used past its expiration date.