JC-1 (CBIC2) is a fluorescent lipophilic carbocyanine dye used to measure mitochondrial membrane potential. JC-1 forms complexes known as J-aggregates at high ΔΨm. Aggregates of JC-1 emit an orange-red fluorescence (Ex/Em=585/590 nm). While in cells with low ΔΨm, JC-1 remains in the monomeric form. JC-1 monomers emit a green fluorescence (Ex/Em=510/527 nm).

Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs).
Labeling of Cells:
1. Culture cells in 6-, 12- , 24-, or 96-well plates at a density of 5× 10⁵ cells/mL. Incubate the cells according to your normal protocol.
2. Ensure that the JC-1 and DMSO has equilibrated to room temperature, and then prepare a 200 μM stock solution by dissolving the contents of one vial in DMSO provided.
3. For the control tube, allow the vial of CCCP has come to room temperature, add 1 μL of CCCP (50 mM). Incubate cells at 37°C for 5 minutes.
4. Add 10 μL JC-1 (200 μM) per well to make the final concentration at 2 μM. Incubate cells at 37°C, 5% CO₂, for 15-20 minutes. If additional labeling followed, for example with an annexin V, begin with step 2.a. If not, proceed with step 1.e.
5. After incubation, centrifuge cells for 3-4 minutes at 400× g at 4°C, carefully aspirate the supernant.
6. Wash cells twice with PBS (1×): add 2 mL PBS (1×) to suspend cells and vortex to mix thoroughly. Centrifuge cells for 3-4 minutes at 400× g at 4°C, carefully aspirate the supernant.
7. Add 500 μL PBS (1×) to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader.

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

652.23

C₂₅H₂₇Cl₄IN₄

3520-43-2

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。
储备液的保存方式和期限：-80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)。-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。

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

  [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.

  [3]. Jung-Ho LEE, In-Hwan LEE, Young-Jun CHOE, 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.