JZL 184 is a potent and selective inhibitor of MAGL with IC50 of 8 nM and 4 μM for inhibition of MAGL and FAAH in mouse brain membranes respectively.

MAGL:8 nM

JZL184是研究内源性2-AG信号效应的有用工具。JZL184展现出对MAGL的时间依赖性抑制作用，并在体外对MAGL比FAAH具有超过300倍的选择性。JZL184不与CB1或CB2受体相互作用，也不抑制2-AG生物合成酶二酰基甘油脂酶-α和二酰基甘油脂酶-β，或花生四烯酸动员酶胞浆磷脂酸A2群IVA。[1]

JZL184在小鼠大脑中迅速且持续地阻断了2-AG水解酶活性，导致内源性2-AG水平升高8倍，并至少维持8小时。接受JZL184处理的小鼠表现出一系列CB1依赖的行为效应，包括镇痛、运动减少和低体温，这表明2-AG介导的内源性大麻素信号在整个哺乳动物神经系统中具有广泛作用。[1]

activity-based protein profiling (ABPP): Mouse brains are Dounce-homogenized in PBS, pH7.5, followed by a low-speed spin (1,400×, 5 min) to remove debris. The supernatant is then subjected to centrifugation (64,000×, 45 min) to provide the cytosolic fraction in the supernatant and the membrane fraction as a pellet. The pellet is washed and resuspended in PBS buffer by sonication. Total protein concentration in each fraction is determined using a protein assay kit. Samples are stored at -80 °C until use. Mouse brain membrane proteomes, are diluted to 1 mg/mL in PBS and pre-incubated with varying concentrations of inhibitors (1 nM to 10 mM) for 30 min at 37 °C before the addition of FP-rhodamine at a final concentration of 2 mM in a 50 mL total reaction volume. After 30 min at 25 °C, the reactions are quenched with 4×SDS-PAGE loading buffer, boiled for 5 min at 90 °C, subjected to SDS-PAGE and visualized in-gel using a flatbed fluorescence s

1 × 105 cells are split into four-well chamber slides and incubated with culture medium containing BrdU for 4 h. BrdU staining is performed following the manufacturer's instructions.(Only for Reference)