Phosphorylation of extracellular signal-regulated kinase as a biomarker for cannabinoid receptor 2 activation
To this end, in this study we evaluated all the downstream signaling of CB2 and developed a method for quantifying CB2R activation in primary mouse and human blood cells based on the findings that (1) CB2R activation results in phosphorylation of extracellular signal-regulated kinase (ERK) in blood cells; and (2) CB2R agonist (e.g. Cell-based quantitative detection of AKT and ERK phosphorylation CHO-K1 cells transiently expressing human or mouse CB2 receptor were seeded in 96-well plates and cultured overnight. 4370), antibody was diluted in the stain/wash buffer (400×) and the cells were incubated in this solution overnight at 4 °C. In the present study, we detected CB2R activation in human primary cells by evaluating AKT and ERK phosphorylation. However, since the pERK signal declines so rapidly following receptor activation, using ERK phosphorylation as a measure of CB2R activation is impractical in vivo.