Human 5-HETE/5-Hydroxyeicosatetraenoic Acid ELISA Kit

A competitive ELISA from Assay Genie designed for the quantitative measurement of 5-HETE (5-hydroxyeicosatetraenoic acid) in human samples. The assay is configured as a Competitive ELISA coated with antigen and is suitable for serum, plasma, cell culture supernatant, cell or tissue lysate, and other liquid samples.

Validated linearity across dilutions of 1:2, 1:4, and 1:8 yields 88–101%, 94–105%, and 87–105% for serum; 82–92%, 83–98%, and 92–99% for EDTA plasma; and 83–98%, 80–97%, and 80–99% for heparin plasma. Recovery averages 97% (88–100%) in serum, 92% (85–99%) in EDTA plasma, and 98% (89–105%) in heparin plasma. The sealed kit should be stored at 2–8°C and not cryopreserved.

The procedure involves adding 50 µL of standard or sample and 50 µL of biotin-labeled antibody per well, mixing, and incubation at 37°C for 45 minutes. After incubation the plate is washed five times, TMB substrate is added and developed in the dark at 37°C for 10–20 minutes, the reaction is stopped (colour changes from blue to yellow), and absorbance is read at 450 nm using a microplate reader.