Mitochondrial Membrane Potential Assay Kit (with JC-1)
中文名称:线粒体膜电位检测试剂盒(JC-1)

Price:
- 应用: 细胞凋亡-JC-1
检测原理 | 线粒体膜电位下降是细胞凋亡早期的标志性事件,发生在细胞核凋亡特征(染色质浓缩、DNA断裂)出现之前,一旦线粒体膜电位崩溃,细胞凋亡便不可逆转。
JC-1是一种广泛用于检测线粒体膜电位 Ψm的理想荧光探针,可以检测细胞、组织或纯化的线粒体膜电位。JC-1有单体和多聚体两种存在形式,两者发射光谱不同。在正常细胞内,线粒体膜电位较高,JC-1聚集在线粒体的基质中,形成聚合物,可产生红色荧光;凋亡早期,线粒体膜电位降低,JC-1不能聚集在线粒体基质中,此时JC-1为单体,可产生绿色荧光。
通过JC-1从红色荧光到绿色荧光的转变可检测到细胞膜电位的下降,可将JC-1荧光颜色的转变作为细胞凋亡早期的检测指标。常用红绿荧光的相对比例来衡量线粒体去极化的比例。
JC-1单体的最大激发波长为514 nm,最大发射波长为529 nm;JC-1聚合物的最大激发波长为585 nm,最大发射波长为590 nm。实际观察时,常规设置红色荧光和绿色荧光即可。 |
检测方法 | 荧光法;流式 |
样本类型 | 细胞 |
检测时间 | 40min |
检测仪器 | 流式细胞仪;荧光显微镜 |
荧光 | JC-1 |
Ex/Em | 514/529, 585/590 |
Channel set | FITC, PE |
自备试剂 | 超纯水 |
保存温度 | 该产品可在2~8°C/-20°C避光条件下保存12个月,具体试剂组分的保存条件请参考说明书。 |
有效期 | 12个月 |
运输条件 | 冰袋 |
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E-CK-A211 | Annexin V-FITC/PI Apoptosis Kit | 200 Assays , 100 Assays , 50 Assays , 20 Assays |
E-CK-A362 | Enhanced Cell Counting Kit 8 (WST-8/CCK8) | 500 Tests , 100 Tests , 500 Tests × 20 |
E-CK-A320 | One-step TUNEL In Situ Apoptosis Kit (Green, FITC) | 100 Assays , 50 Assays , 20 Assays |
Q11:JC-1试剂盒可以将孵育完探针的细胞用1%的多聚甲醛固定,第二天上机么?
不能,染色完后尽量1h内检测上机,若细胞染色完固定,细胞膜的通透性会改变,会影响线粒体膜电位,导致结果不准确。
Q12:JC-1能用荧光酶标仪检测吗?
我们的产品暂时没有在荧光酶标仪上验证过,结果分析方法待开发。
Q13:JC-1成像法的注意事项有哪些?
①显微镜参数一致;②溶解buffer,观察有无红色小颗粒,如果buffer不溶解,可通过12000rpm离心,取上清。③凋亡细胞贴壁能力较弱,会漂浮在培养基中,如果使用荧光显微镜检测,结果相对不准确,建议优先使用流式细胞仪检测;④收集细胞前,拍100x白光照片(小细胞拍200x,比如淋巴细胞)查看形态学变化。
Q14:E-CK-A301线粒体的试剂盒植物组织的样本是否可以用?
植物组织有细胞壁还有叶绿体,可能会对JC-1的检测有影响,目前我们这边没有做过相关验证,不太确定使用效果,如果要做的话,建议参考相关文献并进行预实验
