该步骤仅提供指南，应根据您的特定需求进行修改。在制备ROS Brite™工作溶液之前，根据需要处理细胞。
1）在DMSO中制备10至20mM ROS Brite TM APF（或HPF）储备溶液。通过将DMSO储备溶液稀释到含有20mM Hepes缓冲液（HHBS）的Hanks溶液中，制备1至10μM的工作溶液。
2）根据需要处理细胞（例如，用50-100nM血管紧张素II处理RASM细胞3-5小时）
3）用ROS Brite™APF（或HPF）（1-10μM，步骤＃1）在37℃孵育细胞20-60分钟。
4）用HHBS缓冲液替换染料加载溶液。
5）用适当的荧光仪器在Ex / Em = 490 / 525mm（截止= 515nM）下用底部读取模式分析细胞（例如，用于荧光显微镜的FITC滤光器，用于流式细胞仪的FL1滤光器）。
注意：BSA和酚红会影响荧光，应谨慎使用。 APF和HPF均可用于溶液测定或细胞内测量。
参考文献
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