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.
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JC-1 Mitochondrial Membrane Potential Assay Kit purchased from AbMole
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