The mitochondrial membrane potential was measured according to the protocol of JC-1 Mitochondrial Membrane Potential Assay Kit (Keygen, China). Briefly, the cells were incubated in the incubator for 24h. The cells were collected and washed. JC-1 was added to a final concentration of 1mM with JC-1 monomer (green) as FL1 channel and JC-1 aggregates (red) as FL2 channel.
Jc 1 mitochondrial membrane potential assay kit
Manufactured by Keygen Biotech
Sourced in China
The JC-1 Mitochondrial Membrane Potential Assay Kit is a laboratory instrument designed to measure the mitochondrial membrane potential in cells. It utilizes the fluorescent probe JC-1 to detect changes in the electrochemical gradient across the mitochondrial membrane.
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8 protocols using jc 1 mitochondrial membrane potential assay kit
1
Measuring Mitochondrial Membrane Potential
2
Measurement of Mitochondrial Membrane Potential
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JC-1, a fluorescent lipophilic carbocyanine dye, was used as an indicator by which to measure the mitochondrial membrane potential. MGC80-3 and AGS cells (1.5 × 105 cells/mL) were seeded in 6-well plates. After treatment for 24 hours, AGS and MGC80-3 cells were incubated with 500 μL/well of JC-1 (10 μg/mL) at 37°C for 30 minutes using JC-1 Mitochondrial Membrane Potential Assay Kit (KeyGEN BioTECH, Nanjing, China). The cells were then resuspended twice in 500 μL 1× incubation buffer for 3 minutes each. Finally, they were suspended with 500 μL of 1× incubation buffer. Both red and green fluorescence emissions of mitochondrial membrane potential (Δ ψm) were measured using a flow cytometer (EX = 488 nm, EM = 530 nm).
3
Assessing Mitochondrial Membrane Potential
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Mitochondrial membrane potential was detected using a JC-1 mitochondrial membrane potential assay kit (Nanjing KeyGen Biotech Co., Ltd.), following the manufacturer's protocol. Briefly, after treatment, cells were incubated with JC-1 staining solution at 37°C for 20 min and rinsed twice with incubation buffer provided by the kit. Fluorescence intensity of both mitochondrial JC-1 monomers (Green) and aggregates (Red) were detected using a BD FACSCanto II flow cytometry. JC-1 stains the mitochondria of healthy cells red, and apoptotic cells green.
4
Mitochondrial Membrane Potential Assay
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The JC-1 assay was performed to measure mitochondrial membrane potential (ΔΨm) using the JC-1 mitochondrial membrane potential assay kit (KeyGen, China). Briefly, cells were seeded in a 6-well plate and ΔΨm was detected according to the guidelines of the JC-1 kit. All samples were analyzed using an Accuri or LSRII flow cytometer (BD Biosciences).
5
Mitochondrial Membrane Potential Assay
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Mitochondrial membrane potential (MMP/ΔΨm) assay was performed using the JC-1 Mitochondrial Membrane Potential Assay Kit (KeyGEN, Nanjing, China). JC-1 fluorochrome is a lipophilic cationic mitochondrial vital dye that becomes concentrated in the mitochondrion in response to ΔΨm. The dye exists as a monomer at low concentrations with an emission at 530 nm (green fluorescence, FL1-H), but at higher concentrations, it forms J-aggregates after accumulation in the mitochondrion, with an emission at 590 nm (red fluorescence, FL2-H). To evaluate the ΔΨm for each experimental condition, AR42j cells were seeded in 6-well plates, treated as described above, and washed twice with PBS after experimental treatment; then, 1 mL of staining dye/well (culture medium: JC-1 working dye = 1:1) was added and cells were incubated at 37°C for 20 minutes. Cells were washed twice with cold JC-1 staining buffer and harvested by trypsinization and then examined by flow cytometry.
6
Mitochondrial Potential Evaluation in CRC
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The mitochondrial membrane potential of CRC cells was measured using the JC-1 mitochondrial membrane potential assay kit (Nanjing KeyGen Biotech Co., Ltd.), following the manufacturer’s protocol. In brief, chaetocin treated cells were stained at 37°C for 20 min, then rinsed, using solutions provided in the kit. The fluorescence intensity results were obtained using FACSCanto II flow cytometry (BD Biosciences), whilst mitochondrial JC-1 aggregates which stained red were considered as mitochondria from health cells.
7
Measuring Mitochondrial Membrane Potential
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JC-1 assay was performed to measure mitochondrial membrane potential (ΔΨm) using the JC-1 mitochondrial membrane potential assay kit (KeyGen, China). In short, cells were seeded in 6-well plate and 72 h after siRNA transfection, the detection of ΔΨm was carried out according the guidelines of JC-1 kit. Images were taken by a fluorescent microscope (Nikon, Japan).
8
Mitochondrial Membrane Potential Analysis
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The DCm was analyzed by JC-1 Mitochondrial Membrane Potential Assay Kit (KeyGEN Biotech Inc., China). JC-1 (5, 5′, 6, 6′- tetra-chloro-1,1′,3,3′-tetra-ethylbenzimidazol-carbocyanine iodide) is capable of selectively entering mitochondria, where it forms aggregates and emits red fluorescence when DCm is high. At low DCm, JC-1 cannot enter into mitochondria and forms monomers emitting green fluorescence. The ratio between green and red fluorescence provides an estimate of changes in the mitochondria membrane potential (DCm). MCF-7 cells were treated with desired concentrations of XWL-1-48 for 24 h. After trypsinisation and PBS washing, 1 × 105 cells were incubated for 20 min in freshly prepared JC-1 (1 mM) solution at 37 °C. Spare dye was removed by dye buffer solution washing. The cell-associated fluorescence was measured with FACS (Beckman Coulter, USA).
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