ANNEXIN V- FITC/PI 凋亡检测试剂盒

产品说明：

细胞凋亡是细胞的基本特征之一，在机体的胚胎发育、组织修复、内环境的稳定等方面都起到十分重要

的作用。Annexin V 是一种分子量为35.8 KD 的Ca2+依赖性磷脂结合蛋白，能与细胞凋亡过程中翻转到膜外的

磷脂酰丝氨酸（Phosphatidylserine, PS）高亲和力特异结合。FITC-Annexin V 结合到凋亡细胞后，在蓝色光的

激发下，发出绿色荧光，区分出凋亡细胞与正常细胞。碘化丙啶（Propidium iodide，PI）是一种核酸染料，

它不能穿透完整细胞膜，但对凋亡晚期细胞和死细胞的破损细胞膜能够穿透，并使细胞核红染。将

FITC-Annexin V 与PI 匹配使用，可以将凋亡早期的细胞和晚期的细胞区分开来。

一、产品基本信息

产品名称：ANNEXIN V-FITC/PI凋亡检测试剂盒

货号：CA1020

规格：100T

产品组成成分

Annexin V-FITC 1000μl

Binding Buffer 60ml

Propidium iodide(PI) 1000μl

储存条件：低温

有效期：1年

二、操作步骤

1. 调整待测细胞的浓度为5×105-1×106 个/ml。

2. 取1ml 细胞，1000rpm，4℃离心10 分钟，弃上清。

3. 加入1ml 预冷的PBS，轻轻震荡使细胞悬浮，1000rpm，4℃离心10 分钟，弃上清（对于贴壁细胞，

先用胰酶消化，再用PBS 洗涤）。

4. 重复步骤3 两次。

5. 将细胞重悬于200μl Binding Buffer。

6. 加入10μl Annexin V-FITC 和10μl PI，轻轻混匀，避光室温反应15 分钟 或4℃反应30 分钟。

7. 加入300μl Binding Buffer，在1 小时内用流式细胞仪检测。

8. 若用荧光显微镜检测，将细胞混合液离心，取细胞沉淀涂片，再进行观察。

三、注意事项

为保证得到理想的实验结果在使用此试剂盒之前请认真阅读该注意事项。

1.旋帽离心管装的试剂在开盖前请短暂离心，将盖内壁上的液体甩至管底，避免开盖时液体洒落。

2.Annexin V-FITC 和Propidium iodide 是光敏物质，在保存与操作时请注意避光。

3.Propidium iodide（PI）能通过皮肤吸收，对眼睛有刺激作用。

4.在细胞洗涤的 后一步，请尽量将上清弃净，以免PBS 残留影响实验结果。

5.PI 染色时间过长有可能造成检测的凋亡率偏高，建议首先进行Annexin V-FITC 染色，上机前5 分钟再

加入PI 染色。

6.整个操作过程动作要尽量轻柔，勿用力吹打细胞，尽量在4℃下操作。

7.反应完毕后请尽快检测，因为细胞凋亡是一个动态的过程，反应1 小时后荧光强度就开始衰变。

8.成功的检测凋亡受以下几种因素的影响，如细胞类型、细胞膜上PS 的密度、发生凋亡时PS 翻转的比

例、诱导细胞凋亡的方法、所用试剂、诱导凋亡的时间等，把这些影响因素进行优化对实验成功是非常必要

的。

此试剂盒仅供科研使用。

四、相关产品

P1020 1×PBS，PH7.2-7.4，0.01M

T1300 胰蛋白酶－EDTA 消化液(0.25%)不含酚红

T1350 胰蛋白酶消化液(0.25%)不含酚红

24800 通用细胞冻存液

12100 DMEM(H)

31800 RPMI Medium 1640

CA1040 一步法TUNEL 细胞凋亡检测试剂盒(FITC)

S1056 细胞凋亡荧光Hoechst 33342-PI 双染试剂盒

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