JC-1 Mitochondrial Membrane Potential Assay Kit

JC-1 Mitochondrial Membrane Potential Assay Kit is a kit that uses JC-1 as a fluorescent probe to quickly and sensitively detect the transmembrane potential difference of tissues, cells or purified mitochondria. It can be used for early apoptosis detection.

Components	Size
M9724 JC-1	3 mM in DMSO * 600 uL
M6567 CCCP	50 mM in DMSO * 300 uL

General Protocol

a. 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.

b. 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.

c. Add 5 μL JC-1 (3 mM) per well to make the final concentration at 15 μM. Incubate cells at 37°C, 5% CO2, for 15-20 minutes.

d. After incubation, centrifuge cells for 3-4 minutes at 500×g at 4°C, carefully aspirate the supernant.

e. Wash cells twice with PBS: add 2 mL PBS to suspend cells and vortex to mix thoroughly. Centrifuge cells for 3-4 minutes at 500×g at 4°C, carefully aspirate the supernant.

f. Add 500 μL PBS to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader. (Green fluorescence: Ex/Em=510/527 nm; Red fluorescence: Ex/Em=585/590 nm)

Stored at -20°C protecting from light, and is stable for up to 12 months. Avoid repetitive freeze-thaw cycles while using. For immediate use, components may be stored at 2-8°C.

Precautions

1. A ‘Test’ simply refers to a single assay well. 100 tests based on labeling volumes of 1.0 mL.

2. JC-1 is light sensitive, we recommend that you make small aliquots and store them at -20℃ with light protected.

Product Citations

• 

  Bioact Mater. 2024 Mar 15;37:51-71.

  Microenvironment-responsive metal-phenolic network release platform with ROS scavenging, anti-pyroptosis, and ECM regeneration for intervertebral disc degeneration
  JC-1 Mitochondrial Membrane Potential Assay Kit purchased from AbMole

• 

  Science Advances. 2024 Aug 16;10(33):eado7249.

  Magnetically attracting hydrogel reshapes iron metabolism for tissue repair
  JC-1 Mitochondrial Membrane Potential Assay Kit purchased from AbMole

• 

  Nat Commun. 2023 Aug 23;14(1):5140.

  Self-triggered thermoelectric nanoheterojunction for cancer catalytic and immunotherapy
  JC-1 Mitochondrial Membrane Potential Assay Kit purchased from AbMole

• 

  Research Square. 2021 Jul.

  miR-3587 Targets HMOX1 to Attenuate Ferroptotic Renal Injury Following Ischemia-Reperfusion
  JC-1 Mitochondrial Membrane Potential Assay Kit purchased from AbMole