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The focus of this review is on the qualitative detection of covalent proteinligand adducts but some of the methods enable quantification of time-dependent covalent occupancy, which might be employed to calculate kinetic rate constants reflecting irreversible covalent inhibitor potency [1,60,61]

Protein phosphatase activity assays The protein phosphatase activities were determined using purified proteins from E

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The above procedure was repeated with inhibitor concentrations spanning 0.3 60 M to determine the effective concentration (EC 50 ) required to inhibit 50% NNMT activity in cultured adipocytes

Barcode plasmid pool preparation To generate the bacterial CloneSelect barcode library for the Zeocin resistance marker, a semi-random oligonucleotide pool KI#405 encoding 5-ATGCCGVNNVNNVNNVNNVNNTAA-3 was chemically synthesized (Supplementary Table 2)