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本文介绍一种MTT[3-(4,5二甲基噻唑-2-yl)-2,5-2苯基-四唑溴盐]染色法在测定IFN效价中的应用,结果表明,本法所需指示细胞数少,结果客观重复性好,操作方法简便。 在96孔或40孔细胞培养板上,将对照IFN和待测IFN进行倍比稀释,设细胞和病毒对照。每孔加入100μl 1.5～2×10~5/ml Hep-2细胞悬液,培养5～6小时后,每孔加入100μl 100个TCID_(50)VSV病毒攻击(细胞对照孔不加)。在终止培养前3～4小时,加入5mg/ml MTT 15μl/孔,混匀继续培养,3～4小时后,加入适量生理盐水,用滴管轻轻吹吸,弃去悬浮的病变细胞和死细胞,然后每孔加入200μl
This article describes the application of MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-2-phenyl-tetrazolium bromide] staining method in the determination of IFN titer. Law required to indicate the small number of cells, the results of objective repeatability, the method of operation is simple. On 96-well or 40-well cell culture plates, control IFNs and test IFNs were diluted in multiples, and cell and virus controls were established. Add 100 μl of 1.5-2 × 10-5 / ml Hep-2 cell suspension to each well. After 5 to 6 hours of culture, add 100 μl of 100 TCID50 VSV virus to each well (cell-control wells are not included). 3 to 4 hours before the termination of culture, add 5mg / ml MTT 15μl / well, mix and continue to culture, 3 to 4 hours, add the right amount of saline, gently pipette with a dropper, discarded suspended diseased cells and died Cells were then added 200 [mu] l per well