操作步骤
1.准备HHBS缓冲液
2.在高质量无水DMSO中制备2 mM至5 mM Mag-Fluo-4 AM储备液
2.1使用的Mag-Fluo-4 AM量：1毫克
2.2所需浓度：2 mM
2.3在合适的容器中，将1mg Mag-Fluo-4AM与611.51μL无水DMSO混合。
3.使用10μMMag-Fluo-4 AM 2在HHBS中制备2X工作溶液
3.1终孔内浓度的Mag-Fluo-4 AM：5μM
3.2在合适的容器中混合16μL的Mag-Fluo-4AM。接下来，添加HHBS或您选择的缓冲液，直到体积为3.2 mL。
注意：对于大多数细胞系，我们建议Mag-Fluo-4 AM的终浓度为4至5μM。
4.将100μL染料工作溶液加入已经含有100μL培养基的所需孔中。
4.1该步骤将染料工作溶液从2X稀释至1X，并将每种组分的终浓度调节至5μM的Mag-Fluo-4 AM
5.孵育染料
5.1将染料加载板在细胞培养箱中孵育30-60分钟。
5.2将染料加载板在室温下孵育30分钟。
6.准备HHBS缓冲液（或您选择的缓冲液）。
6.1在合适的容器中加入HHBS或您选择的缓冲液，直至体积为4 mL。
7.用HHBS缓冲液或您选择的缓冲液替换染料工作溶液，使用1.0 mM Probenecid。
7.1首先，从所需孔中除去200μL染料工作溶液和培养基。
7.2在相同的孔中加入200μL含有1.0mM丙磺舒的HHBS（或您选择的缓冲液）。
8.运行实验
8.1为您的样品添加所需的处理。
8.2以Ex / Em = 494/516 nm运行实验。
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