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JC-1 Mitochondrial Membrane Potential Assay Kit

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Based on 165 publication(s) in Google Scholar

MCE JC-1 Mitochondrial Membrane Potential Assay Kit uses JC-1 to detect the mitochondrial membrane potential in variety of cell types, as well as intact tissues and isolated mitochondria.

  • Storage :

    Stored at -20°C protecting from light, and is stable for up to 1 year. Avoid repetitive freeze-thaw cycles while using. For immediate use, components may be stored at 2-8°C.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • JC-1 Mitochondrial Membrane Potential Assay Kit Product Image
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Description & Advantages

ΔΨm, mitochondrial membrane potential, is an important parameter of mitochondrial function and has been used as an indicator of cell health. Variation of ΔΨm would be studied using JC-1. In healthy cells with high ΔΨm, JC-1 forms complexes known as J-aggregates. While in cells with low ΔΨm, JC-1 remains in the monomeric form. When excited at 510 nm, JC-1 monomers emit a green fluorescence with a maximum at ~527 nm. Aggregates of JC-1 emit an orange-red fluorescence with a maximum at ~590 nm.

MCE JC-1 Mitochondrial Membrane Potential Assay Kit uses JC-1, a lipophilic cationic dye, to detect the mitochondrial membrane potential in variety of cell types, as well as intact tissues and isolated mitochondria.

MCE JC-1 Mitochondrial Membrane Potential Assay Kit consists of JC-1, Phosphate-buffered saline (10×) and CCCP (50 mM in DMSO). CCCP controls should be used to confirm that the JC-1 response is sensitive to changes in membrane potential.

MCE JC-1 Mitochondrial Membrane Potential Assay Kit can be analyzed by fluorescence microscopy, flow cytometer, or a fluorescence plate reader with appropriate filter sets.

Protocol

1. Preparation of Phosphate-buffered saline (PBS) (1×)

a. Warm the PBS (10×) until any salt crystals are completely dissolved.

b. Dilute PBS (10×) 1:10 with ddH2O.

2. Labeling of Cells

a. Culture cells in 6-, 12- , 24-, or 96-well plates at a density of 5-10× 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℃ for 5 minutes.

c. Add 10 μL JC-1 (200 μM) per well to make the final concentration at 2 μM. Incubate cells at 37℃, 5% CO2, for 15-20 minutes.

d. After incubation, centrifuge cells for 3-4 minutes at 400× g at 4℃, carefully aspirate the supernant.

e. 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℃, carefully aspirate the supernant.

f. Add 500 μL PBS (1×) to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader.

3. Additional Labeling (using Annexin V as an example, not provided in this kit)

a. After step 2.c, 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℃, carefully aspirate the supernant.

b. Add 100 μL Annexin V binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4) to suspend the JC-1-stained cells.

c. Add appropriate volume of Annexin V and incubate cells at 37℃, 5% CO2, for 15 minutes.

d. Add 400 μL Annexin V binding buffer (10 mM HEPES, 140 mM NaCl, 2.5 mM CaCl2, pH 7.4) to suspend cells. Analyze sample on a flow cytometer, fluorescence microscopy, or fluorescence microplate reader.

Storage

Stored at -20°C protecting from light, and is stable for up to 1 year. Avoid repetitive freeze-thaw cycles while using. For immediate use, components may be stored at 2-8°C.

Components

Components JC-1 (200 μM in DMSO) Phosphate-buffered saline (10×) CCCP (50 mM in DMSO)
HY-K0601-100T 230 μL × 5 25 mL 125 μL

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  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Nat Nanotechnol. 2025 Jul;20(7):935-946. [Abstract]

    Representative CLSM images of the mitochondrial membrane potential (2 μM JC-1; 37℃ for 20 min) of CT26 cells after different treatments. Scale bar, 10 μm.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Apr 5;16(1):3253. [Abstract]

    Analysis of mitochondrial membrane potential in Hep G2 and MDA-MB-231 cells treated with 100 nm Au@MIL NPs for 24, 48, and 72 h using JC-1 staining. Enlarged cells (white arrows) show JC-1 fluorescence localized in the cytoplasm. Mitochondrial depolarization increased over time, as indicated by the rising green signal.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2025 Nov 18:e12793. [Abstract]

    Flow cytometry analysis of mitochondrial membrane potential using JC‐1 staining in mice Ti‐Tregs treated with PBS or PUE (2.0 mg kg−1day−1). The ratio of JC‐1 aggregates (red) to monomers (green) was calculated (n = 6).

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Signal Transduct Target Ther. 2023 Aug 16;8(1):302. [Abstract]

    Flow cytometry results indicating the alternative mitochondrial membrane potential induced by 8 mM or 16 mM D-lactate. JC-1 (2 μM; 37 ℃ for 20 min) monomers and JC-1 aggregates represent low and high MMP, respectively.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: J Neuroinflammation. 2023 Jan 30;20(1):16. [Abstract]

    Representative graphs show JC-1 fuorescence of microglia in vitro and quantitative analysis of green/red fuorescence intensity (%), red fuorescence represents the mitochondrial aggregate JC-1 and green fuorescence indicates the monomeric JC-1.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Nano Energy. 2022: 108002.

    JC-1 staining of 4T1 cells as a measure of mitochondrial depolarization after different therapies. JC-1 aggregate indicates mitochondria with normal membrane potential (red), and JC-1 monomer means the mitochondria with a depolarized and damaged membrane (green) (Scale bar = 40 µm).

Assay Types
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
    1/16

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Nat Nanotechnol. 2025 Jul;20(7):935-946. [Abstract]

    Representative CLSM images of the mitochondrial membrane potential (2 μM JC-1; 37℃ for 20 min) of CT26 cells after different treatments. Scale bar, 10 μm.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
    1/16

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Nat Commun. 2025 Apr 5;16(1):3253. [Abstract]

    Analysis of mitochondrial membrane potential in Hep G2 and MDA-MB-231 cells treated with 100 nm Au@MIL NPs for 24, 48, and 72 h using JC-1 staining. Enlarged cells (white arrows) show JC-1 fluorescence localized in the cytoplasm. Mitochondrial depolarization increased over time, as indicated by the rising green signal.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
    1/16

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Adv Sci (Weinh). 2025 Nov 18:e12793. [Abstract]

    Flow cytometry analysis of mitochondrial membrane potential using JC‐1 staining in mice Ti‐Tregs treated with PBS or PUE (2.0 mg kg−1day−1). The ratio of JC‐1 aggregates (red) to monomers (green) was calculated (n = 6).

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
    1/16

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Signal Transduct Target Ther. 2023 Aug 16;8(1):302. [Abstract]

    Flow cytometry results indicating the alternative mitochondrial membrane potential induced by 8 mM or 16 mM D-lactate. JC-1 (2 μM; 37 ℃ for 20 min) monomers and JC-1 aggregates represent low and high MMP, respectively.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
    1/16

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: J Neuroinflammation. 2023 Jan 30;20(1):16. [Abstract]

    Representative graphs show JC-1 fuorescence of microglia in vitro and quantitative analysis of green/red fuorescence intensity (%), red fuorescence represents the mitochondrial aggregate JC-1 and green fuorescence indicates the monomeric JC-1.

  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
    1/16

    JC-1 Mitochondrial Membrane Potential Assay Kit purchased from MedChemExpress. Usage Cited in: Nano Energy. 2022: 108002.

    JC-1 staining of 4T1 cells as a measure of mitochondrial depolarization after different therapies. JC-1 aggregate indicates mitochondria with normal membrane potential (red), and JC-1 monomer means the mitochondria with a depolarized and damaged membrane (green) (Scale bar = 40 µm).

  • JC-1 Mitochondrial Membrane Potential Assay Kit Product Image
    1/16
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • Experimental Validation Results for JC-1 Mitochondrial Membrane Potential Assay Kit
  • JC-1 Mitochondrial Membrane Potential Assay Kit Product Image