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MTT试剂盒/细胞毒性检测试剂盒/细胞增殖检测试剂盒


相关信息

英文名称 MTT Cell Proliferation and Cytotoxicity Assay Kit
储存条件 -20℃,避光,有效期1年
单位
规格    500T

产品简介:
MTT可以被线粒体内的一些脱氢酶还原生成结晶状的深紫色产物formazan,在特定溶剂存在的情况下,可以被完全溶解,然后通过酶标仪可以测定490nm波长附近的吸光度。细胞增殖越多越快,则吸光度越高;细胞毒性越大,则吸光度越低。
使用说明:
1.收集对数期细胞,调整细胞悬液浓度,分于96孔板,每孔180μL,3000-10000个细胞/孔。
2.置37℃、5%CO2温箱培养使细胞贴壁,培养6-24小时。
3.加入适当浓度的受试化合物,继续培养适当时间。
4.小心吸去上清,加入90μL新鲜培养液,再加入10μL MTT溶液,继续培养4 h。
5.然后吸掉上清,每孔加入110μL Formazan溶解液,置摇床上低速振荡10 min,使结晶物充分溶解。在酶联免疫检测仪490 nm处测量各孔的吸光值。
6.同时设置调零孔(培养基、MTT、 Formazan溶解液),对照孔(细胞、相同浓度的药物溶解介质、培养液、MTT、 Formazan溶解液),每组设定3复孔。
注意事项:
1.由于使用96孔板进行检测,如果细胞培养时间较长,一定要注意蒸发的问题。一方面,由于96孔板周围一圈最容易蒸发,可以采取弃用周围一圈的办法,改加PBS,水或培养液;另一方面,可以把96孔板置于靠近培养箱内水源的地方,以缓解蒸发。
2.MTT溶液为黄色需避光保存,长时间光照会导致失效。当颜色变为灰绿色时,请勿使用。Formazan溶解液结冻或产生沉淀时可以37℃水浴孵育以促进溶解,并且必须在全部溶解并混匀后使用。
3.为了您的安全和健康,请穿实验服并戴一次性手套操作。



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