Human Kinesin Family, Member 1A ELISA Kit

This sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of human KIF1A in tissue homogenates or other biological fluids. From Antibodies.com, the kit uses a pre-coated 96-well microplate format with an antibody specific for KIF1A immobilised on the plate wells. Standards and samples are added and KIF1A present is bound by the immobilised antibody. Following incubation and washing, Biotinylated Anti-KIF1A Antibody is added, which binds the captured KIF1A, followed by HRP-Avidin conjugate. TMB substrate solution is then used to visualise the HRP enzymatic reaction; the catalysed product is blue and changes to yellow upon addition of acidic stop solution. The density of yellow is proportional to the amount of KIF1A captured, and concentration is calculated by reading the absorbance at 450 nm in a colorimetric microplate reader and referring to a standard curve.

The quantitative assay has a reported range of 0.156–10 ng/ml and a sensitivity of <0.051 ng/ml. Total assay time is 4 hours 30 minutes. Components are stored at +4°C or −20°C as instructed, and the kit should not be used past its expiration date.

Sample recovery has been characterised for serum, EDTA plasma and heparin plasma. Reported average values are 91% for serum (n=5, range 83–100%), 96% for EDTA plasma (n=5, range 90–103%) and 95% for heparin plasma (n=5, range 89–102%). Dilution linearity has been assessed at 1:2, 1:4, 1:8 and 1:16: serum (n=5) at 83–110%, 79–100%, 90–98% and 82–108%; EDTA plasma (n=5) at 90–98%, 90–105%, 73–109% and 87–91%; heparin plasma (n=5) at 88–103%, 77–105%, 95–98% and 87–110%.