Cell Meter 细胞凋亡活性检测试剂盒*红色荧光*

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流式细胞仪
Ex:	488 nm
Em：	575/26  nm
通道:	PE 通道

样品实验方案

简要概述

1.根据需要处理样品。
2.准备Xite β-D-吡喃半乳糖苷工作溶液并将其添加到样品中。
3.在37°C下孵育样品15至45分钟。
4.使用Cy3/TRITC滤光片组监控荧光强度。

 

溶液制备 

1.储备溶液

所有未使用的储备溶液应分为一次性使用的等分试样，并在制备后储存在-20°C下。 避免重复冻融循环。
Xite β-D-吡喃半乳糖苷储备溶液（100X）：
将100 uL DMSO（组分C）添加到Xite β-D-吡喃半乳糖苷（组分A）中并充分混合。 注意：将未使用的Xite β-D-吡喃半乳糖苷储备溶液以单次使用的等分试样保存在-20°C下。

 

2.工作溶液

Xite β-D-吡喃半乳糖苷工作溶液（1X）：
用1 mL测定缓冲液稀释10 uL Xite β-D-吡喃半乳糖苷储备溶液（100X），制成Xite β-D-吡喃半乳糖苷工作溶液（1X）。注意：应立即使用Xite β-D-吡喃半乳糖苷工作溶液。

 

样品操作及实验分析

以下方案可用作指导，具体实验应根据需要进行调整。

1.根据需要处理样品。

2.用您选择的缓冲液（例如DPBS）洗涤细胞。

3.加入100 uL Xite β-D-吡喃半乳糖苷工作溶液15-45分钟，然后在37°C的培养箱中孵育样品。 注意：孵育的时间需要仔细确定。

4.取出工作溶液并用您选择的缓冲液洗涤细胞。

5.将细胞重悬于测定缓冲液（组分B）中，并使用流式细胞仪或带有FITC过滤器的荧光显微镜监控荧光强度。

 

参考文献

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Stochastic modeling of aging cells reveals how damage accumulation, repair, and cell-division asymmetry affect clonal senescence and population fitness
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Topological DNA damage, telomere attrition and T cell senescence during chronic viral infections
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Leucine reduces the proliferation of MC3T3-E1 cells through DNA damage and cell senescence
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Regulatory T cells trigger effector T cell DNA damage and senescence caused by metabolic competition
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Hydrogen Treatment Protects against Cell Death and Senescence Induced by Oxidative Damage
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MicroRNA-16 feedback loop with p53 and Wip1 can regulate cell fate determination between apoptosis and senescence in DNA damage response
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