Coelenterazine h is a dehydroxylated derivative of native coelenterazine and serves as a substrate for Renilla luciferase (Rluc) as well as a cofactor for jellyfish luminescent protein. Its luminescence intensity is more than 10 times higher than that of native coelenterazine, making it suitable for reporter gene analysis. Additionally, it can be used for the detection of intracellular calcium ion concentrations, gene reporter analysis, bioluminescence resonance energy transfer (BRET) studies, ELISA, high-throughput screening (HTS), and chemiluminescent detection of reactive oxygen species (ROS) levels in tissues or cells.