XTT细胞活力分析试剂盒是一种比色分析方法,可以检测细胞的代谢能力。在分析方法中,黄色四氮唑在代谢活跃的细胞中可以变成高度着色的甲臜染料。这种转变只存在于活力旺盛的细胞中,因此甲臜染料的产生量与样本中活力旺盛的细胞成正比。在此方法中形成的甲臜染料溶于水,可以用分光光度计在波长为450nm处测其吸光值进行定量。电子偶联剂如PMS( N-甲基吩嗪 甲基硫酸盐)可以显著提高XTT的降低效率。XTT细胞活力分析试剂盒可以检测到在细胞中线粒体酶作用下由XTT转变成的甲臜染料。 因为线粒体酶在细胞死亡后失去活性,橘红色的甲臜染料只存在活力旺盛的细胞中。 XTT细胞活力分析试剂盒有望在大部分细胞系中应用。细胞类型和孵育时间的不同, 0.2-2x10 4 cells/well对于大多数实验室足够的。为了取得更理想的结果,建议做细胞浓度梯度细胞活力和增殖分析广泛应用于药物的研发,以进一步研究生长因子、细胞因子和细胞毒制剂。在早期药物发现的复合筛选和后期药品的安全性和毒性研究中,高通量筛查需要可靠、敏感和简单的分析方法,以具备分析大量样本的能力。采用四氮唑如MTT、XTT、WST-1等比色法分析细胞活力的方法是在活细胞中四氮唑变成高度着色的甲臜染料复合物基础上发展的(1、2)。相比之下,细胞增殖分析方法如活细胞中用放射性碘和BrdU标记的DNA后,用合并碘(对样本的放射性定量)或BrdU(用anti-BrdU 抗体)来定量,而XTT分析方法不需要放射性物质、固定细胞或细胞透化。因此,与上述细胞分析方法不同,用XTT分析过的细胞还可以用于进一步分析。 1000 assays (96 well format). 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. 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.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.