For typical assay, dissolve enzyme at a concentration of 1 mg/ml in 0,05 M TES with 0,36 mM calcium chloride, pH 7,5. Dilutions run are 1/10 and 1/20 in the above buffer.

One unit liberates one µmole of L-leucine equivalents from collagen in 5 hours at 37 °C, pH 7,5.

Activators: Ca²⁺ is required; Inhibitors: Metal chelating agents such as cysteine, EDTA or o-phenanthroline but not DFP. It is also inhibited by a2-macroglobulin.