JC-1 Mitochondrial Membrane Potential Assay Kit JC-1线粒体膜电位检测试剂盒

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Overview

JC-1 Mitochondrial Membrane Potential Assay Kit JC-1线粒体膜电位检测试剂盒

Specifications

Brand翌圣生物 Yeasen

Cat.No40706ES

Specification100 T

Origin翌圣生物科技(上海)股份有限公司

技术平台 荧光检测

应用方法 细胞凋亡研究检测早期凋亡与线粒体膜电位变化 线粒体功能研究评估线粒体膜电位及去极化程度

检测方法 荧光探针法 红/绿荧光比值法

生物特性参数 试剂类型线粒体膜电位检测试剂盒 检测靶标线粒体膜电位(ΔΨm) 作用机制JC-1电势依赖性聚集；膜电位降低时由红色聚合物转为绿色单体 检测类型定性和定量检测 荧光探针JC-1 应用早期细胞凋亡检测 检测指标红/绿荧光强度比值反映线粒体去极化程度 相关生物过程细胞凋亡早期线粒体膜通透性改变

样本类型细胞组织 纯化的线粒体

技术参数 储存条件-20℃避光保存，避免反复冻融，有效期一年 运输条件冰袋运输 阳性对照CCCP 检测样本数六孔板100个样品；12孔板200个样品 荧光波长聚合物 Ex=585 nm/Em=590 nm；单体 Ex=514 nm/Em=529 nm 缓冲液储存条件超纯水和JC-1染色缓冲液(5×)可4℃保存 有效期一年

实验类型 线粒体膜电位检测通过JC-1红绿荧光变化检测ΔΨm 细胞凋亡检测用于早期细胞凋亡检测

FAQ

Q：JC-1 探针和试剂盒有什么区别，客户做细胞实验用哪一款？

A:前者只是一个探针的粉末；后者是液体，又多了缓冲液，阳性对照和超纯水。两个都可以。

Q：JC-1 试剂盒检测的时候，客户要用流式检测，是要先把贴壁细胞消化下来再按照悬浮细胞处理吗？还是先染完色再消化下来去检测？

A:先消化下来制成细胞悬液再染色。

Q：阴性对照中线粒体都是正常的线粒体，为什么也可以观察到绿色的荧光，而且红色的也看不到细胞形态的原因是？

A：阴性对照可以拍到绿色可能是因为染色的时候加入的染料较多，从而使阴性对照可以排除绿色的荧光，可以适当的降低探针的浓度。绿色可以看到整个形态是因为单体（绿光）是散落在细胞质中的，而聚合体（红色）是结合到线粒体上的，有些细胞中线粒体分布是不均匀的，能否完整的看到细胞的形态是和线粒体的分布也是有一定的原因的。

Q：石蜡切片和冰冻切片可以做JC-1吗？

A：不可以，JC-1实验的样本必须是活细胞。

Q：JC-1孵育完成后不能及时检测，可以固定后再检测吗？

A：不可以，JC-1用于活细胞样本的检测，固定后细胞死亡；此外，染色后放置过久会导致荧光淬灭，建议染色30 min内完成检测。

Q：组织样本可不可以用JC-1检测凋亡？

A：无法直接检测组织，可先将组织制备成单细胞悬液，再按照悬浮细胞的操作步骤来检测，需对制备方案进行摸索。

Q：JC-1工作液的配置中出现绿色沉淀应如何处理？

A：取适量JC-1（200×），按照每50μL JC-1（200×）加入8 mL 超纯水的比例稀释JC-1，剧烈震荡充分溶解并混匀JC-1，然后再加入2 mL JC-1 染色缓冲液（5×），如溶液后存在绿色不溶沉淀可加入千分之一的Tween 20。

Q：该JC-1是否可用于分离纯化的完整线粒体膜电位的检测？JC-1的建议工作液如何配置？对应的纯化线粒体的量应为多少？加入JC-1后多久可以检测？

A：可以用于纯化分离得到的完整线粒体的膜电位的检测。

JC-1的工作液及线粒体用量的比例为：0.9 mL JC-1染色工作液（1×）中加入0.1 mL总蛋白量为10-100 µg纯化的线粒体。

加入JC-1工作液3-5 min后即可进行检测。

但需留意对于冷冻的纯化线粒体不可用于进行膜电位的检测，因线粒体膜经冻融被破坏。

Documents & Citations

细胞功能检测和探针整体方案 Open document

Published citations

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