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Competitive ABPP study

AD Achintya Kumar Dolui
AV Arun Kumar Vijayakumar
RR Ram Rajasekharan
PV Panneerselvam Vijayaraj
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To determine the sensitivity of the detected serine hydrolases towards well-known inhibitors of SHs, we performed the competitive ABPP labeling with known serine hydrolase inhibitors such as PMSF, Paraoxon, and Profenofos. Competitive serine hydrolase labeling was conducted with 100 µg of soluble protein by pre-incubation with either 100 μM inhibitor or DMSO (as a control) for 30 min at 37 °C. After incubation, 2 μM of the probe was added into each reaction mixture and kept for 1 h at 37 °C in the dark. Separation and detection of ABPP labeled proteins were done as described above.

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