噻唑兰(Methylthiazolyldiphenyl-tetrazolium bromide, MTT),也称作溴化噻唑蓝四氮唑,是一种黄色染料,已经普遍替代传统的台盼蓝染色法或者放射性同位素插入法,用来检测细胞的活力,细胞增殖以及细胞毒性分析。检测原理在于:MTT是一种可接受氢离子的化合物染料,带正电荷,具有细胞膜渗透性。活细胞线粒体中的琥珀酸脱氢酶能够将外源进入的MTT还原成为水不溶性的深蓝色MTT-甲臜结晶,而死细胞不具有此功能。甲臜结晶是不能穿透细胞膜,因此沉积在处于增殖且未受损伤的细胞内。甲臜结晶可用DMSO或者酸化的异丙醇溶解出来,酶标仪下测定570 nm的吸光度反映甲臜的生成量。而甲臜的生成量与活细胞数目成正比,用以评估细胞存活状况。
本试剂盒以MTT(高纯度)作为指示剂,在经典操作步骤的基础上,采用独特的甲臜溶解液配方,可以直接溶解甲臜沉淀,无需去除原有的培养液。从而避免因去除培养液时甲臜被部分去除而引起的操作误差。具有本底低,灵敏度高,线性范围宽,操作简单等特点。另外,本试剂盒还提供配套的一次性针头过滤器和注射器,为实验带来更多的便利。
冰袋运输。
收到产品后,取出除菌套装室温保存,其他组分4℃保存。甲臜溶解液也可以放到室温保存。一年有效。
Q:MTT 配置成工作液后的使用注意事项?
A:MTT 配制成溶液后为黄色,需避光保存,长时间光照会导致失效。当颜色变为灰绿色时, 请勿使用。MTT 溶液在 4℃、冰浴等较低温度情况下会凝固,可以 20-25℃水浴温育片刻至全部融解 后使用。
Q:配好的 MTT 溶液是否需要过滤?
A:无需过滤。
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