Recombinant human acetylcholinesterase

Recombinant human acetylcholinesterase (hAChE), Torpedo californica acetylcholinesterase (tcAChE), acetylthiocholine iodide (ATCI), 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB), all-trans-retinoic acid, thiazolyl blue tetrazolium bromide (MTT), isopropanol, dimethyl sulfoxide (DMSO), and the drugs donepezil hydrochloride, MW = 415.95 g/mol; tacrine hydrochloride, MW = 234.72; berberine hydrochloride, MW = 371.82 g/mol; and galantamine hydrobromide, MW = 368.27 g/mol, were all purchased from Sigma-Aldrich (Poole, UK). Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), albumin, and poly-D-lysine (PDL) were purchased from Thermofisher Scientific (Rochester, UK). Unless specified differently, all other chemicals were from Sigma-Aldrich (Poole, UK).

Profenofos, 4-bromo-2-chlorophenol (BCP), bovine serum albumin (BSA), reduced nicotinamide adenine dinucleotide phosphate (NADPH), 5,5-dithiobis (2-nitrobenzoic acid) (DTNB) and acetylthiocholine iodide (ATC) were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands). Magnesium chloride hexahydrate (MgCl₂⋅6H₂O), trifluoroacetic acid (TFA), dimethylsulfoxide (DMSO), and calcium chloride dihydrate (CaCl₂⋅2H₂O) were purchased from VWR International (Amsterdam, The Netherlands). Acetonitrile (ACN, UPLC/MS grade) was purchased from Biosolve (Valkenswaard, The Netherlands). Human liver microsomes and cytosol (pooled from 150 human donors, mixed gender), rat liver microsomes and rat liver cytosol (pooled from 20 male Sprague–Dawley rats) were purchased from Corning (Amsterdam, The Netherlands). Recombinant human acetylcholinesterase was purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands). Rat blood was purchased from BioIVT (Westbury, USA), human and rat plasma were purchased from Innovate (New York, USA).