Carboxylesterase 2 (CES2) is a member of serine esterase family which can hydrolyze ester and amide bonds. The protein Involved in the detoxification of xenobiotics and in the activation of ester and amide prodrugs. Hydrolyzes aromatic and aliphatic esters, but has no catalytic activity toward amides or a fatty acyl-CoA ester. Hydrolyzes the methyl ester group of cocaine to form benzoylecgonine. Thus, the recombinant human CES2 activity was measured by its ability to hydrolyze 4-Nitrophenyl acetate (4-NPA) to 4-Nitrophenol. The reaction was performed in 50 mM Tris, 150 mM NaCl, pH 7.5 (Assay Buffer), ainitiated by addition 50 μL of various concentrations of CES2 (diluted by Assay Buffer) to 50 μL of 2 mM Substrate 4-NPA (100 mM stock in Acetone, diluted by Assay Buffer). Incubated at 37℃ for 10 min, then read at a wavelength of 400 nm.