产品描述:
本试剂盒利用荧光探针DHE进行活性氧检测。DHE可自由透过活细胞膜进入细胞内,在细胞质中呈蓝色荧光,被细胞内的ROS氧化后形成氧化乙啶掺入染色体DNA中,使细胞核呈现明亮的红色荧光。在激发波长535nm,发射波长610nm附近,使用荧光显微镜、激光共聚焦显微镜、荧光分光光度计、荧光酶标仪、流式细胞仪等检测荧光强度,测定细胞内活性氧水平。具有背景低、灵敏度高、线性范围宽、使用方便等优点,是一种快速简便的组织或培养活细胞中ROS经典检测方法。
适用范围:
贴壁细胞、悬浮细胞、新鲜动物组织
工作波长:
激发波长480-535nm,最佳发射波长590-610nm
产品组成:
0.2ml, 5mM DHE
储存条件:
-20ºC避光保存,一年有效
所需设备:
流式细胞仪、荧光酶标仪、激光共聚焦显微镜、荧光分光光度计等。
注意事项:
(1) 开盖前请短暂离心,将盖内壁上的液体收集于管底,避免开盖时染液损失;
(2) 要设有无DHE孵育的对照,即不加探针,只用0.01M PBS重悬的细胞;
(3) 荧光探针标记后,一定要洗净残余的未进入细胞内的探针,以避免背景升高。多次清洗时注意操作规范,避免将细胞洗掉;
(4) 对于药物处理(孵育)时间较短的细胞(2小时以内、也有建议4小时以内),可以先用荧光染料进行标记,后用药物刺激细胞后。对于药物处理时间较长的细胞(超过4小时或6小时以上),建议先用药物处理(刺激)细胞,后用探针标记;
(5) DHE在光照和空气中易被氧化,保存和操作中应注意避光;
(6) 对不同的细胞和组织,应选择合适的孵育时间和浓度,以观察ROS的变化;
(7) 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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