Measuring Mast Cell Degranulation

BMMCs were cultured with rIL-3 and rSCF in the presence or absence of 500 nM TSA for 1 h or overnight. Cells were activated and supernatants and cell lysates were collected 1 h later. β-hexosaminidase (β-hex) activity was assessed, as previously described (28 (link), 59 (link)). Briefly, cells were washed and supernatants and pellets were collected. Pellets were lysed with 0.5% Triton X-100. Both supernatants and pellets were then treated with 4-nitrophenyl-N-acetyl-β-D-glucosaminide (p-NAG) substrate (Sigma) for 1 h. Plates were read at 405 nm using a spectrophotometer to determine β-hexosaminidase activity. Data are depicted as percent specific release according to the following formula: (Stimulated supernatants/(supernatant ± pellet)^*100−unstimulated supernatants/(supernatant ± pellet)^*100).