MMP was measured using the JC-1 assay (Beyotime). Cells were observed under a fluorescence microscope (Nikon, Tokyo, Japan). The fluorescence intensity was analyzed by the software ImageJ (NIH, Bethesda, MD, USA), and the ratio of red to green fluorescence reflected the MMP [37 (link)].
Jc 1 assay
Manufactured by Beyotime
Sourced in China
The JC-1 assay is a fluorescent probe used to measure mitochondrial membrane potential in cells. It can detect changes in the electrochemical gradient across the mitochondrial inner membrane, which is an indicator of mitochondrial function and cell health.
Automatically generated - may contain errors
Lab products found in correlation
Fluorescence microscope, by Nikon (1 mentions)
Lcs sp8 sted, by Leica (1 mentions)
Mitosox red, by Thermo Fisher Scientific (1 mentions)
C6 flow cytometry, by BD (1 mentions)
Atp assay kit, by Nanjing Jiancheng (1 mentions)
Confocal laser scanning microscope, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Eclipse ti microscope, by Nikon (1 mentions)
Ferroorange, by Dojindo Laboratories (1 mentions)
13 protocols using jc 1 assay
1
Mitochondrial Membrane Potential Measurement
2
Mitochondrial Membrane Potential Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial membrane potential was measured using the JC-1 assay (Beyotime). Cells were observed under a confocal microscope (Leica-LCS-SP8-STED, Leica, Germany). The fluorescence intensity was analyzed by the software ImageJ (NIH, Bethesda, MD, USA), and the ratio of red to green fluorescence reflected the MMP.
3
Mitochondrial Oxidative Stress Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
L02 and BEAS-2B cells were
seeded into six-well glass-bottom plates and treated with NPs at the
same concentrations as those used in the TEM observations (L02 with
0, 0.0125, and 0.125 mg/mL; BEAS-2B with 0, 0.0125, and 0.25 mg/mL).
The medium was removed after exposure, and 5 μM MitoSOX Red
(M3600, Thermo) was added to each well. The cells were then incubated
for 10 min, subsequently washed with PBS, and then fixed with 4% PFA
for 20 min. The MMP was determined using a 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetra-ethylbenzimidazolocarbo-cyanine
iodide (JC-1) assay (Beyotime Institute of Biotechnology, Beijing,
China) according to the manufacturer’s instructions. An equal
volume (1 mL) of 5 μg/mL JC-1 staining solution was added to
the cells that were then incubated for 20 min and subsequently washed
with PBS. Carbonyl cyanide m-chlorophenyl hydrazone
was used as a positive control. The presence of mROS and MMP in cells
was observed by CLSM. The excitation and emission wavelengths were
as follows: MitoSOX Red, 510 and 580 nm; mitochondrial JC-1 monomers,
∼490 and 530 nm; and JC-1 aggregates, ∼525 and 590 nm,
respectively.
seeded into six-well glass-bottom plates and treated with NPs at the
same concentrations as those used in the TEM observations (L02 with
0, 0.0125, and 0.125 mg/mL; BEAS-2B with 0, 0.0125, and 0.25 mg/mL).
The medium was removed after exposure, and 5 μM MitoSOX Red
(M3600, Thermo) was added to each well. The cells were then incubated
for 10 min, subsequently washed with PBS, and then fixed with 4% PFA
for 20 min. The MMP was determined using a 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetra-ethylbenzimidazolocarbo-cyanine
iodide (JC-1) assay (Beyotime Institute of Biotechnology, Beijing,
China) according to the manufacturer’s instructions. An equal
volume (1 mL) of 5 μg/mL JC-1 staining solution was added to
the cells that were then incubated for 20 min and subsequently washed
with PBS. Carbonyl cyanide m-chlorophenyl hydrazone
was used as a positive control. The presence of mROS and MMP in cells
was observed by CLSM. The excitation and emission wavelengths were
as follows: MitoSOX Red, 510 and 580 nm; mitochondrial JC-1 monomers,
∼490 and 530 nm; and JC-1 aggregates, ∼525 and 590 nm,
respectively.
4
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial membrane potential (MMP) was assessed using the JC-1 assay (Beyotime, C2006, China). Following the instructions, the JC-1 working solution was prepared. After the cell collection from each group, they were thoroughly mixed with 500 μL of the JC-1 working solution, and incubated at 37°C for 20 minutes. Then, the cells were washed twice with precooled 1× JC-1 assay Buffer and then tested by a C6 flow cytometry (Becton Dickinson, USA).
5
Measuring Mitochondrial Function and ATP in CRC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mitochondrial membrane potential was measured by JC-1 assay (Beyotime). HCT116 and DLD-1 cells were incubated in a mixture of culture medium and JC-1 working solution in a 1:1 ratio in the dark for 20 min at 37 °C. Then, the cells were washed twice to remove free JC-1. The fresh medium was changed before the acquisition of images under a laser scanning confocal microscope (Olympus, Japan).
The ATP content of the CRC cells was measured using an ATP assay kit (Nanjing Jiancheng Bioengineering Institute, China) by colorimetry of phosphomolybdic acids according to the manufacturer’s instructions.
The ATP content of the CRC cells was measured using an ATP assay kit (Nanjing Jiancheng Bioengineering Institute, China) by colorimetry of phosphomolybdic acids according to the manufacturer’s instructions.
6
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mitochondrial membrane potential (MMP) was determined using a JC-1 assay (Beyotime Institute of Biotechnology, China) according to the manufacturer’s instructions. PC9 or PC9GR cells were plated in 6-well plates (2.0 × 106 cells/well) and incubated for 24 h, then cultured with different concentrations of apatinib for another 24 h. After incubation, cells were harvested with trypsin, washed twice with PBS, supplemented with 500 μL JC-1 dye staining solution, and then incubated at 37 °C for 30 min in the dark. After incubation, the cells were centrifuged at 400g × 5 min and washed twice with 1× incubation buffer. The cells were then resuspended in 500 μL 1× incubation buffer and fluorescence was detected using flow cytometry (488 nm excitation and 525 nm emission filters).
7
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial membrane potential was assessed by JC‐1 assay (Beyotime). JC‐1 is a membrane‐permeable dye that selectively enters mitochondria; change in mitochondrial membrane potential can be detected by the fluorescence transition of aggregates. Hep3B cells were treated with 10 mg/ml HP for 0, 6, and 12 hr prior to incubation with 20 mM JC‐1 for 15 min at 37°C and then washed with PBS. Nuclei were visualized by DAPI (Solarbio life sciences) staining. After washing with PBS, cells were imaged using a fluorescence core cell culture microscope (EVOS®xl core cell culture microscope); fluorescence intensity was assessed qualitatively.
8
Mitochondrial Membrane Potential Measurement
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mitochondrial membrane potential was measured by JC-1 assay (Beyotime, C2003S). HUVECs were incubated in a mixture of culture medium and JC-1 working solution for 30 min at 37 ℃. Then, the cells were washed 3 times to remove the free JC-1 reagent. After changed with the fresh medium, the images were captured under a Nikon ECLIPSE Ti microscope (Nikon, Japan).
9
Macrophage Mitochondrial and Iron Assays
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Macrophages were planted in 48-well plates and treated according to group. Mitochondria membrane potential (JC-1 assay, Beyotime, C2006) and iron ion (ferroOrange, DOJINDO, F374) staining were performed on macrophages according to manufacturer's instructions.
10
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mitochondrial membrane potential (ΔΨm) of the cells was measured using the JC-1 assay (Beyotime). BPH-1 cells were seeded in six-well plates (3 × 105 cells/well) and incubated at 37°C for 24 h. Next, the cells were treated with different concentrations of TMJ-12 (0, 8, and 16 µM) for 48 h, after which, the cells were collected and incubated with 10 mM JC-1 in the dark at 37°C for 30 min. Finally, JC-1 fluorescence was measured using the flurescence microscope (BD Biosciences, NJ, U.S.A.).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!