反应详情

3T3 Swiss Albino fibroblast cell suspension (Phenol-red free DMEM with 10% FBS and 2 mM L-glutamine) was plated out into 96-well tissue culture plates (1×105 cells/mL, 100 μL/well). The cells were cultured overnight at 37° C., 5% CO2. The extracts and fractions were solubilised in DMSO, and diluted appropriately with media so the extracts were tested at a final concentration of 1000, 10, and 1 μg/mL; 5 μL of sample was added to each well. Cells+samples were incubated (37° C., 3 hours), before the addition of calcium ionophore A23187. Following 20 min incubation, the plate was centrifuged (1000 g, 3 min) and the supernatants were removed. Included on the plate was a positive control (aspirin; 100 μM), and DMSO control, both with and without calcium ionophore A23187.

1. additiondiluted appropriately with media so the extracts
2. addition5 μL of sample was added to each well
3. custom(37° C., 3 hours)
4. waitFollowing 20 min
5. custom(1000 g, 3 min)
6. customthe supernatants were removed