JC-1 (solution) 

JC-1 (CBIC2) (solution) is an ideal fluorescent probe widely used to detect mitochondrial membrane potential. JC-1 accumulates in mitochondria in a potential dependent manner and can be used to detect the membrane potential of cells, tissues or purified mitochondria. In normal mitochondria, JC-1 aggregates in the mitochondrial matrix to form a polymer, which emits strong red fluorescence (Ex=585 nm, Em=590 nm) ; When the mitochondrial membrane potential is low, JC-1 cannot aggregate in the matrix of mitochondria and produce green fluorescence (ex=510 nm, em= 527 nm) ^[1].
Solvent and concentration: DMSO: 1.5 mM

JC-1 工作液配制
用预热好的无血清细胞培养基或 PBS 稀释储存液，配制成 1.5-30 μM 的 JC-1 工作液。
注：1) 请根据实际情况调整 JC-1 工作液浓度，且现用现配。
2) 如果 JC-1 进入细胞的效果不好，可向工作也液中加入适量 20% Pluronic F127 溶液，终浓度为 0.02-0.05%，Pluronic F127 可以防止 JC-1 在缓冲液中聚合并帮助其进入细胞。
JC-1染色
1) 以 6 孔板为例进行细胞铺板，密度为 5×10⁵ cells/mL。37°C，5% CO₂ 培养箱培养过夜。
注：进行凋亡诱导时的细胞密度建议不超过 1×10⁶/mL，也可根据自己的细胞类型培养至合适的密度。
2) 取 0.5 mL 细胞悬液至无菌的离心管内。
3) 400 g 离心 3-5 min；弃上清。
4) 用 1 mL JC-1 工作液重悬细胞，于 37°C，5% CO₂ 培养箱孵育 15-30 min。
5) 室温条件 400 g 离心 5 min；吸掉上清。
6) 用 2 mL 细胞培养液或者缓冲液重悬细胞，之后室温条件 400 g 离心 5 min；弃上清，重复两次。
7) 用 1 mL 新鲜培养液或者缓冲液重悬细胞，立刻进行后续的流式分析或荧光显微镜观察。
8) 数据分析 (流式细胞仪) ：含有红色 JC-1 聚集物的健康细胞线粒体用 FL2 通道检测；含有绿色 JC-1 单体的凋亡或不健康细胞用 FL1 (FITC) 通道检测。
注：若用于酶标仪检测，用 300 μL 缓冲液重悬细胞；然后按照每孔 100 μL 的量将染色好的细胞转移到不透光的 96 孔板内，即可进行荧光酶标板分析。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

652.22

C₂₅H₂₇Cl₄IN₄

3520-43-2

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. A Perelman, et al. JC-1: alternative excitation wavelengths facilitate mitochondrial membrane potential cytometry. Cell Death Dis. 2012 Nov 22;3:e430.  [Content Brief]

  [2]. Vera C. Keil, et al. Ratiometric high-resolution imaging of JC-1 fluorescence reveals the subcellular heterogeneity of astrocytic mitochondria. Pflügers Archiv - European Journal of Physiology. 2011,462(5): 693-708.  [Content Brief]

  [3]. Jung-Ho LEE, et al. Real-time analysis of amyloid fibril formation of α-synuclein using a fibrillation-state-specific fluorescent probe of JC-1. Biochem. J. 2009, 418:311-323.

  [4]. Salvioli S, et al. JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: implications for studies on mitochondrial functionality during apoptosis. FEBS Lett. 1997 Jul 7;411(1):77-82.  [Content Brief]