活性氧 Cell Meter 荧光法胞内总ROS检测试剂盒 适合于流式细胞仪
| Ex (nm) | - | Em (nm) | - |
| 分子量 | - | 溶剂 | - |
| 存储条件 | - |
活性氧（ROS）是氧正常代谢的天然副产物，在细胞信号传导中起重要作用。但是，在与氧化应激相关的状态下，ROS水平会急剧增加。 ROS的积累会严重破坏细胞结构。氧化应激在心血管疾病，糖尿病，骨质疏松症，中风，炎性疾病，许多神经退行性疾病和癌症中的作用已得到公认。 ROS检测将有助于确定氧化应激如何调节各种细胞内途径。 Cell Meter 荧光法胞内总ROS检测试剂盒使用我们独特的Amplite ROS Green荧光定量活细胞中的ROS，Amplite ROS Green具有细胞渗透性。与ROS反应时会生成绿色荧光。 Cell Meter 荧光法胞内总ROS检测试剂盒提供了一种灵敏的一步荧光测定法，可在培养一小时后检测活细胞中的细胞内ROS。该试剂盒针对流式细胞仪应用进行了优化，其信号可以通过Ex / Em = 490/520 nm（FL1通道）进行检测。百萤生物是AAT Bioquest的中国代理商，为您提供优质的Cell Meter 荧光法胞内总ROS检测试剂盒。
活性氧（ROS）篇：包含总ROS和多种活性氧离子检测试剂大全
适用仪器
| 流式细胞仪 | |
| Ex: | 488 nm |
| Em： | 530/30 nm |
| 通道: | FITC 通道 |
样品实验方案
简要概述
1.准备密度为0.5-1×106细胞/ mL的细胞
2.在0.5 mL细胞悬液中加入1 µL 500X Amplite ROS Green
3.在37℃下将细胞染色1小时
4.处理细胞以诱导ROS
5.使用带有FL1通道的流式细胞仪分析细胞（Ex / Em = 490/520 nm）
溶液配制
1.储备溶液配制
除非另有说明，否则所有未使用的储备溶液应分成一次性等分试样，并在制备后储存在-20°C。 避免反复冻融循环。
Amplite ROS绿色储备溶液（500X）：将100 µL DMSO（组分C）添加到Amplite ROS Green（组分A）的小瓶中，并充分混合以制成500X Amplite ROS Green储备液。 避光。 注意：要存放，请密封管。有关细胞样品制备的指南，请点击查看。
操作步骤
1.对于每个样品，以0.5×105至1×106细胞/ mL的密度在0.5 mL测定缓冲液（组分B）或自备缓冲液中制备细胞。注意：应单独评估每个细胞系，以确定诱导ROS的细胞密度。
2.将1 µL 500X Amplite ROS Green储备溶液加入0.5 mL细胞悬液中。
3.在37ºC下孵育1小时。注意：对于贴壁细胞，用0.5 mM EDTA轻轻提起细胞以保持细胞完整，并在与Amplite ROS Green孵育之前用含血清的培养基洗涤细胞一次。合适的孵育时间取决于所用的单个细胞类型和测试化合物，优化每个实验的孵育时间。
4.通过在所需的缓冲液（例如PBS或HBSS）中添加50 µL 11X测试化合物来处理细胞。对于对照孔（未处的细胞），添加相应量的缓冲液。
5.将细胞在37ºC下孵育，以诱导ROS（避光）。注意：我们在37℃下用100 µM TBHP（氢过氧化叔丁基）处理Jurkat细胞30分钟，以诱导ROS。有关详细信息，请参见图1。
6.使用具有FL1通道（Ex / Em = 490/520 nm）的流式细胞仪检测荧光强度。
图示
| 图1.使用Cell Meter 荧光法细胞内总ROS活性测定试剂盒在TBHP处理后检测Jurkat细胞中的细胞内ROS。 将细胞与Amplite ROS Green在37°C孵育1小时。 然后将细胞在不使用（蓝色）或（红色）100 µM TBHP的情况下在37°C下处理30分钟。 使用流式细胞仪（BD FACSCalibur）在FL1通道处检测荧光信号。 |
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