Human Apolipoprotein F ELISA Kit

This sandwich ELISA kit from Antibodies.com enables the in vitro quantitative determination of human Apolipoprotein F (APOF) in serum, plasma or other biological fluids. The assay provides a sensitivity of 0.066 ng/ml and a quantitative range of 0.156–10 ng/ml, run on a pre-coated microplate configured as 12 x 8 well strips and read at 450 nm on a colorimetric microplate reader. Total assay time is 4h 30m, and kit components are stored at +4°C or -20°C as instructed.

Spike-and-recovery performance was assessed in three matrices. Serum (n=5) gave 80–102% recovery with an average of 91%; EDTA plasma (n=5) gave 81–100% with an average of 90%; and heparin plasma (n=5) gave 80–89% with an average of 84%. Dilution linearity at 1:2, 1:4, 1:8 and 1:16 was 87–91%, 87–107%, 74–101% and 92–97% for serum (n=5); 90–105%, 84–101%, 90–101% and 79–108% for EDTA plasma (n=5); and 84–95%, 92–105%, 82–105% and 89–91% for heparin plasma (n=5).

The assay uses a sandwich enzyme immunoassay format: an APOF-specific antibody is pre-coated on the plate, standards or samples are added, and the captured APOF is detected with a Biotinylated Anti-APOF Antibody followed by HRP-Avidin conjugate. TMB substrate develops a blue product that turns yellow on addition of stop solution, and APOF concentration is calculated from the O.D. absorbance at 450 nm against the standard curve.