手机验证
22901
Fluorimetric Intracellular Total ROS Activity Assay Kit*Red Fluorescence*
红色
活性氧胞内总ROS检测试剂盒 红色荧光
Fluorimetric Intracellular Total ROS Activity Assay Kit*Red Fluorescence*
活性氧胞内总ROS检测试剂盒 红色荧光
200T
适用仪器
荧光显微镜 | |
Ex: | 520 nm |
Em: | 605 nm |
推荐孔板: | 黑色透明底板 |
通道: | Texas Red 通道 |
荧光酶标仪 | |
Ex: | 520 nm |
Em: | 605 nm |
Cutoff: | 590 nm |
推荐孔板: | 黑色透明底板 |
读取模式: | 底部读取 |
样品实验方案
简要概述
溶液配制
储备溶液配制
1. Amplite ROS Red储备液(500X):将40 µL DMSO(组分C)添加到Amplite ROS Red(组分A)的小瓶中,并充分混合以制成500X Amplite ROS Red储备液。 避光。 注意:20 µL 500X Amplite ROS Red储备液足以用于1个板。 注意:如果将试管紧密密封并避免光照,可以将未使用的部分等分并在<-20°C下保存超过一个月。 避免重复冻融循环。
工作溶液配制
将20 µL 500X Amplite ROS Red储备溶液添加到10 mL的测定缓冲液(组分B)中,并充分混合以制成Amplite ROS Red工作溶液。 注意:此Amplite ROS Red工作溶液在室温下至少可稳定2小时。
实验步骤
1.将100 µL /孔(96孔板)或25 µL /孔(384孔板)的Amplite ROS Red工作溶液添加到细胞板中。
2.将细胞在5%CO2、37°C的培养箱中孵育一小时。
3.在所需的缓冲液(例如PBS或HHBS)中,用20µL 11X测试化合物(96孔板)或10 µL 6X测试化合物(384孔板)处理细胞。 对于对照孔(未处理的细胞),添加相应量的化合物缓冲液。
4.要诱导ROS,请在室温下或在5%CO2、37°C的培养箱中孵育细胞板至少15分钟(对于用1 mM H2O2处理的Hela细胞为30分钟)。
5.在Ex / Em = 520/605 nm(截止= 590 nm)处使用荧光酶标仪(底部读取模式)检测荧光的增加,或在Ex / Em = 520/605 nm滤光片组(Texas Red)下使用荧光显微镜观察细胞)。
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