Optimal activity is seen in its unique buffer. NEBuffer 2 gives around 50% activity. Optimal activity is seen at a Mg²⁺ concentration of 3 mM. At 10 mM Mg²⁺, activity drops to 60%. The salt optimum is tight around 75 mM (1).

(1) Gerard G. F., and D' Alessio J. M., Chapter 6 (73-93) From: Methods in Molecular Biology, Vol.16: Enzymes of Molecular Biology Edited by: M. M. Burell 1993 Humana Press Inc. Totowa, NJ