We describe a sensitive and reproducible lipase assay based on the binding of 63Ni to fatty acid. This method can detect down to 1 nmol of fatty acid per milliliter of solution. It has been adapted for measuring low concentrations of lipoprotein lipase and hepatic triacylglycerol lipase. Furthermore, in the presence of tritiated triolein, the method is insensitive to radiolabel interference, even when the fatty acid is labeled.