活性氧 Cell Meter 荧光法胞内总ROS检测试剂盒 红色荧光
| 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测量将有助于确定氧化应激如何调节各种细胞内途径。 Amplite 荧光ROS检测试剂盒使用我们独特的ROS指示剂来定量活细胞中的ROS。 Amplite ROS Red具有细胞渗透性。与ROS反应时会产生红色荧光。该试剂盒是一种优化的“混合和读取”测定形式。 Amplite 荧光ROS测定试剂盒提供了一种灵敏的一步荧光测定法，可在1-2小时的孵育中检测活细胞中的细胞内ROS。该测定可以以方便的96孔或384孔板形式进行，无需分离步骤即可轻松实现自动化。使用荧光酶标仪或荧光显微镜可以轻松读取其信号。它可用于定量ROS活性或筛选ROS抑制剂。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
| 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
- 在生长培养基中准备细胞
- 加入Amplite ROS Red工作溶液（对于96孔板为100 µL /孔，对于384孔板为25 µL /孔）
- 在37°C下孵育细胞1小时
- 用测试化合物处理细胞以诱导ROS
- 在Ex / Em = 520/605 nm（截止= 590 nm）或安装Ex / Em = 520/605 nm滤光片的荧光显微镜下检测荧光增加（底部读取模式）
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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