Cell Meter Annexin V凋亡检测试剂盒 深红色荧光 适合流式细胞检测
Ex (nm) | 656 | Em (nm) | 670 |
分子量 | - | 溶剂 | - |
存储条件 | - |
我们的Cell Meter 检测试剂盒是一套用于检测细胞生存力的工具,有多种参数可用于监测细胞活力。 该特定试剂盒旨在通过测量磷脂酰丝氨酸(PS)的转运来检测细胞凋亡。 在细胞凋亡中,PS转移到质膜的外部小叶。 磷脂酰丝氨酸在细胞表面的出现是细胞凋亡初始/中间阶段的普遍指标,可以在观察形态变化之前进行检测。 该试剂盒使用与PS特异性结合的荧光膜联蛋白V。 已经证明膜联蛋白V缀合物选择性结合PS。 使用带有Cy5滤波片的流式细胞仪优化了该特定的检测试剂盒,以检测细胞凋亡。百萤生物是AAT Bioquest的中国代理商,为您提供优质的Cell Meter Annexin V凋亡检测试剂盒。
适用仪器
流式细胞仪 | |
Ex: | 640 nm |
Em: | 660/20 nm |
通道: | APC 通道 |
样品实验方案
简要概述
- 用测试化合物制备细胞(200 µL /样品)
- 添加膜联蛋白V-iFluor 647测定溶液
- 在室温下孵育30-60分钟
- 使用带有660/20 nm滤光片(APC通道)的流式细胞仪或带有Cy5滤光片组的荧光显微镜分析细胞
实验步骤
1.用膜联蛋白V-iFluor 647制备和孵育细胞:
1.1用测试化合物处理细胞一段时间(对于用星形孢菌素处理过的Jurkat细胞,需要4-6小时)以诱导细胞凋亡。
1.2离心细胞以获得1-5×105细胞/管。
1.3将细胞重悬于200 µL分析缓冲液(组分B)中。
1.4向细胞中加入2 µL Annexin V-iFluor 647(组分A)。
1.5避光保存,于室温下孵育30至60分钟。
1.6在使用流式细胞仪或荧光显微镜分析细胞之前,添加300 µL分析缓冲液(组分B)以增加体积。
1.7使用带有660/20 nm滤光片的流式细胞仪(APC通道)或带有Cy5滤光片组的荧光显微镜检测荧光强度。
2.通过使用流式细胞仪进行分析:
2.1使用带有660/20 nm滤光片(APC通道)的流式细胞仪定量膜联蛋白V-iFluor 647的结合物。
3.通过使用荧光显微镜进行分析:
3.1孵育后用移液管吸移细胞,用测定缓冲液冲洗1-2次,然后用测定缓冲液重悬细胞。
3.2将细胞添加到载玻片盖玻片的载玻片上。 注意:对于贴壁细胞,建议直接在盖玻片上生长细胞。 与Annexin V-iFluor 647一起孵育后,用测定缓冲液冲洗1-2次,然后将测定缓冲液加回到盖玻片上。 将玻片上的盖玻片倒置并可视化细胞。 与膜联蛋白V-iFluor 647孵育后,还可以将细胞固定在2%甲醛中,并在显微镜下观察。
3.3使用Cy5滤光片组,在荧光显微镜下用Annexin V-iFluor 647分析凋亡细胞。当碘化丙锭添加到细胞中时,通过使用TRITC通道测量细胞活力。质膜上的橙色染色表明膜联蛋白V-iFluor 647与细胞表面的PS结合。
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