HCID 21584101, compound → 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide

HRID 1397195Notes

Reaction equation

HRID 1397195 reaction equation
[Br-:1].[CH3:2][C:3]1[N:4]=[C:5]([N:9]2[N:13]=[C:12]([C:14]3[CH:19]=[CH:18][CH:17]=[CH:16][CH:15]=3)[NH2+:11][N:10]2[C:20]2[CH:25]=[CH:24][CH:23]=[CH:22][CH:21]=2)[S:6][C:7]=1[CH3:8].N=NC=NN>C(=O)=O>[CH3:2][C:3]1[N:4]=[C:5]([N+:9]2[N:10]([C:20]3[CH:25]=[CH:24][CH:23]=[CH:22][CH:21]=3)[N:11]=[C:12]([C:14]3[CH:19]=[CH:18][CH:17]=[CH:16][CH:15]=3)[N:13]=2)[S:6][C:7]=1[CH3:8].[Br-:1]

Participants

4

Reactant 2

HCID 21584101HCID 21584101, compoundHCID 53427792HCID 53427792, compound2×

Product 1

HCID 649653-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide

Reagents, catalysts & solvents 1

HCID 280Carbon DioxideSolvent

Procedure

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.

Workup

  1. waitthe cells were incubated with 5 mg/mL MTT reagent for four-hours
  2. customThe formazan precipitates
  3. dissolutionwere then dissolved by addition of 0.1 mL of HCl in isopropanol
  4. additionmixed thoroughly

Sources & evidence

ORD record
ord-35b1f22438644e2aa25c66bcdf22cfc7
Source dataset
uspto-grants-2014_12
Patent number
US08901168B2

Reaction notes

Loading…