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CAS RN: | Product Number: J0043
JC-1 Mitochondrial Membrane Potential Assay Kit
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Product Information
- Components : JC-1 dye 100 nmol×1vial, CCCP solution (10mM in DMSO) 500 μL×1vial
Description
- JC-1 is widely used in the observation of mitochondrial membrane potential, as it shows a characteristic fluorescence shift from green (530 nm) to red (590 nm) depending on the presence of a mitochondrial membrane potential. Mitochondrial membrane potential is an important facet of mitochondrial function and has been used as an indicator of cell health. In normal mitochondria, JC-1 collects and aggregates within the mitochondria. This aggregate fluoresces red. However, when the mitochondrial membrane potential is low, such as after having been depolarized, JC-1 cannot collect and aggregate within the mitochondria, leaving it as monomers with a green fluorescence. In this way, mitochondrial depolarization can be tracked through the red/green fluorescence intensity ratio, with smaller ratios indicating a larger extent of depolarization. To allow for comparison with depolarized cells as a control, this kit includes the depolarizing agent CCCP in a DMSO solution. JC-1 aggregates can be detected with filter sets designed to detect rhodamine (λex/λem = 540/570 nm). JC-1 monomers can be detected with filter sets designed to detect FITC (λex/λem = 485/535 nm).
Direction for Use
- Preparation of JC-1 DMSO stock solution (2mM)
- Add 50μL of DMSO to the included tube of JC-1 Dye and dissolve by pipetting.
- * Use prepared JC-1 DMSO stock solution within 1 month.
- Preparation of JC-1 working solution
- First, add an appropriate amount of the JC-1/DMSO stock solution to the bottom of a microtube. Add an appropriate amount of medium and mix by pipetting. Adjust so the final concentration of JC-1 is in the range of 1-15 μM.
- * If the fluorescence signals are weak, optimization of JC-1 concentration and/or incubation time may be necessary.
- *When preparing JC-1 working solution, it is imperative that medium be added to the JC-1/DMSO stock solution. Reversing the order greatly increases the chances of JC-1 precipitating.
- *Use the JC-1 working solution within 1 day.
- Preparation of CCCP working solution
- Add an appropriate amount of CCCP/DMSO stock solution (10mM) into a microtube. Add an appropriate amount of medium and mix by pipetting. Adjust the final concentration of CCCP to fall within the range of 5-100 µM.
- * If mitochondrial membrane depolarization is not observed, optimize the concentration of CCCP in working solution and/or incubation time.
- * Use the CCCP working solution within 1 day.
- Mitochondrial membrane potential determination using JC-1
- Seed cells into a dish or a chamber slide and incubate at 37°C in a 5% CO2 incubator.
- Prepare a control sample by applying CCCP. Incubate the cells at 37°C in a 5% CO2 incubator for 30–90 minutes.
- Remove half of the medium from each well (both samples and CCCP controls). Add to each well an equivalent amount of 2x JC-1 working solution.
* For example, if the final concentration of JC-1 is to be 2 μM, apply a 4 μM JC-1 working solution to wells. - Incubate the cells at 37°C in a 5% CO2 incubator for 20–60 minutes.
- Remove medium from wells, and wash 3x with PBS.
- After the final wash, observe the cells under a fluorescence microscope, or alternatively, measure fluorescence intensity via a plate reader.
Storage
- Store at -20°C. This product is photosensitive and should be protected from light. Use within 6 months.
Product Number | J0043 |
Physical State (20 deg.C) | Liquid |
Storage Temperature ![]() |
Frozen (-20°C) |
Condition to Avoid | Heat Sensitive |
Specifications
Properties (reference)
GHS
Signal Word | Warning |
Hazard Statements | H227 : Combustible liquid. |
Precautionary Statements | P501 : Dispose of contents/ container to an approved waste disposal plant. P210 : Keep away from heat, hot surfaces, sparks, open flames and other ignition sources. No smoking. P280 : Wear protective gloves/ eye protection/ face protection. P370 + P378 : In case of fire: Use dry sand, dry chemical or alcohol-resistant foam to extinguish. P403 : Store in a well-ventilated place. |
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Transport Information:
H.S.code* | 3822.19-000 |
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