使用细胞活力指标AM酯类
AM酯是非极性酯，其易于穿过活细胞膜，并且通过活细胞内的细胞酯酶快速水解。 AM酯广泛用于非侵入性地将各种极性荧光探针加载到活细胞中。 但是，使用AM酯时必须小心，因为它们易于水解，特别是在溶液中。 它们应在使用前重新配制成高质量的无水二甲基亚砜（DMSO）。 DMSO储备溶液可以在20℃下干燥储存并避光。 在这些条件下，AM酯应稳定数月。
以下是我们推荐的将AM酯加载到活细胞中的方案。 该说明书仅提供指南，实际应根据您的具体需求进行修改。
a）在高质量无水DMSO中制备2至5mM AM酯的储备溶液。非离子洗涤剂Pluronic®F-127有时用于增加AM酯的水溶性。
注意：在稀释到上样缓冲液中之前，可将等体积的20％Pluronic®F-127溶液添加到细胞活力指示剂的DMSO储备溶液中。终的Pluronic®F-127浓度约为0.02％。
注意：不建议在Pluronic®F-127存在下长期储存AM酯。
b）在实验当天，要么将细胞活力的固体指示剂溶解在DMSO中，要么将等分的指示剂储备溶液解冻至室温。在您选择的缓冲液（如Hanks和Hepes缓冲液）中制备1至10μM的工作溶液。对于大多数细胞系，建议终浓度范围为4至5μM的细胞活力指标。细胞加载所需指标的确切浓度必须根据经验确定。
c）如果您的细胞含有有机阴离子转运蛋白，可以在细胞培养基中加入丙磺舒（1-2.5 mM）或磺吡酮（0.1-0.25 mM），以减少脱酯化指标的泄漏。在室温或37℃下将细胞与AM酯孵育20分钟至1小时。
d）在不含细胞活力指示剂的缓冲液（含有阴离子转运蛋白抑制剂，如果适用）中洗涤细胞以除去过量的探针。
注意：通过流式细胞术立即分析细胞样品，以确定在零时间没有洗涤的每个细胞的平均荧光。
e）在所需的Ex / Em波长下进行实验 。
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相关产品
| 产品名称 |
货号 |
| 钙黄绿素, AM |
Cat#22002 |
| 钙黄绿素红,AM |
Cat#21900 |
| 钙红素醋酸盐 |
Cat#22010 |
