| 货号 | 9095S |
| 描述 | The XTT Cell Viability Assay Kit is a colorimetric assay that detects the cellular metabolic activities. During the assay, the yellow tetrazolium salt XTT is reduced to a highly colored formazan dye by dehydrogenase enzymes in metabolically active cells. This conversion only occurs in viable cells and thus, the amount of the formazan produced is proportional to viable cells in the sample. The formazan dye formed in the assay is soluble in aqueous solution and can be quantified by measuring the absorbance at wavelength 450 nm using a spectrophotometer. An electron coupling reagent, such as PMS (N-Methylphenazonium methyl sulfate), can significantly improve the efficiency of XTT reduction in cells. |
| 应用 | FUNC |
| 目标/特异性 | The XTT Cell Viability Kit detects formazan dye produced from XTT conversion by mitochondrial enzymes in cells. Because these mitochondrial enzymes are inactivated shortly after cell death, the orange colored formazan dye only appears in viable cells. This XTT Cell Viability Kit is expected to work in most cells lines. Variable with cell type and incubation time employed in the assay, 0.2-2x104 cells/well should be sufficient for most experiments. For the best result, a cell number titration (as shown in Figure 1) is recommended. |
| 供应商 | CST |
| 背景 | Cell viability and proliferation assays are widely used in drug discovery for the study of growth factors, cytokines, and cytotoxic agents. High throughput screening, in early drug discovery compound screening and in later drug safety and toxicity studies, requires a reliable, sensitive, and simple assay with the ability to analyze a large number of samples. Colorimetric cell viability assays using tetrazolium salt, such as MTT, XTT, WST-1, etc. were developed based on live cells reduction of tetrazolium salt into highly colored formazan compounds (1,2). In contrast to cell proliferation assays, such as radioactive thymidine or BrdU labeling of DNA in live cells followed by quantification of the incorporated thymidine (by quantifying sample radioactivity) or BrdU (using anti-BrdU antibody), the XTT assay doesn’t require radioactive materials, cell fixing, or cell permeabilization. Thus, unlike alternative cellular analysis assays, cells examined in the XTT assay may be used for further analysis. |
| 存放说明 | -20C |
| 参考文献 | 1 . Scudiero, D.A. et al. (1988) Cancer Res 48, 4827-33. 2 . Roehm, N.W. et al. (1991) J Immunol Methods 142, 257-65. |
Figure 1. C2C12 cells were seeded at varying density in a 96-well plate and incubated overnight. The XTT assay solution was added to the plate and cells were incubated. The absorbance at 450 nm was measured at 1.0, 2.0, 3.0, 4.0, and 5.0 hours.C2C12细胞按照不同的密度种于96孔板中，孵育过夜。将XTT分析液加到板中然后孵育细胞，在450nm处，分别于1.0、2.0、3.0、4.0和5.0小时测吸光值。 | |
Figure 2. C2C12 cells were seeded at 2x104 cells/well in a 96-well plate and incubated overnight. Cells were then treated with various concentrations of doxorubicin overnight. The cytotoxicity was measured using XTT Cell Viability Kit (red) and BrdU Cell Proliferation Assay Kit #6813 (blue) as shown in the left panel. The percentage inhibition in each assay was calculated and plotted in the right panel. Doxorubicin treatment can lead to cell DNA damage followed by cell cycle arrest.C2C12细胞按照2x10 4 cells/well 种于96孔板中孵育过夜。然后用不同浓度的链霉素处理细胞过夜。用XTT细胞活力分析试剂盒（红色）和BrdUp细胞增殖分析试剂盒（蓝色）检测细胞毒性，如左图所示。右图中显示在每个分析方法中的抑制比例。链霉素可以引起细胞DNA的损伤，进而使细胞周期停止。 | |
Figure 3. HeLa cells were seeded at 1x104 cells/well in a 96-well plate and incubated overnight. Cells were then treated with various concentrations of staurosporine overnight. The cytotoxicity was measured using XTT Cell Viability Kit (red) and BrdU Cell Proliferation Assay Kit #6813 (blue) as shown in the left panel. The percentage inhibition in each assay was calculated and plotted in the right panel. Staurosporine is a nonspecific kinase inhibitor and induces cellular apoptosis.HeLa细胞按照1x10 4 cells/well种于96孔板中孵育过夜。然后用不同浓度的十字孢碱处理细胞过夜。用XTT细胞活力分析试剂盒（红色）和BrdUp细胞增殖分析试剂盒（蓝色）检测细胞毒性，如左图所示。右图中显示在每个分析方法中的抑制比例。十字孢碱是一种非特异性激酶抑制剂，可以引起细胞凋亡。 |
