HCID 21584101, compound → (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)

Participants

Reaction Steps

Proliferation of splenocytes was measured using the MTT assay (Hansen et al., 1989), which measures the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide to formazan crystals. In this assay, the yellow MTT solution will be converted to a purple formazan product by the mitochondrial dehydrogenase of the viable cells. One-hundred microliters of splenocytes (1×107 cells/mL) were seeded in triplicates in flat-bottomed 96-well plates and cultured in the presence or absence of 1 μg/mL Con A or LPS or 10 μg/mL TT. After 72 hours at 37° C. in 5% CO2, the cells were incubated with 5 mg/mL MTT reagent for four-hours. The formazan precipitates were then dissolved by addition of 0.1 mL of HCl in isopropanol and mixed thoroughly by repeated pipetting. Cell proliferation was quantified using an ELISA reader at 570 nm.

Provenance & Evidence