适用仪器

样品实验方案

简要概述

1. 用5×10⁵至1×10⁶细胞/ mL的密度制备含测试化合物的细胞
2. 将1 µL 500X Nuclear Red CCS2添加到0.5 mL细胞溶液中
3. 在37°C，5％CO₂下孵育10-30分钟
4. 使用带有PE-Texas Red通道的流式细胞仪分析细胞（Ex / Em = 488 nm / 615 nm）

实验步骤

1.对于每个样品，以0.5×10⁵至1×10⁶细胞/ mL的密度在0.5 mL温暖的培养基或自备缓冲液中制备细胞。注意：应单独评估每个细胞系，以确定最佳细胞密度。

2.用测试化合物处理细胞一段时间，以诱导细胞凋亡，细胞周期停滞或其他细胞周期功能。

3.在含有生长培养基的细胞中加入1 µL 500X Nuclear Red CCS2（组分A）。

4.将细胞在37°C，5％CO₂培养箱中孵育10至30分钟。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，并在用Nuclear Red CCS2孵育之前用含血清的培养基洗涤一次。合适的孵育时间取决于所用的单个细胞类型和细胞浓度。优化每个实验的孵育时间。染色前无需固定细胞，因为Nuclear Red CCS2具有细胞渗透性。

5.可选：将细胞以1000 rpm的速度离心4分钟，然后将细胞重悬于0.5 mL的测定缓冲液（组分B）或自备的缓冲液中。

6.使用带有PE-Texas Red通道（Ex / Em = 488/615 nm）的流式细胞仪检测荧光强度。

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