Cellular mitochondrial dysfunction can be reflected by the loss of the mitochondrial membrane potential, which can be indirectly measured by the fluorescent probe JC-1. JC-1 is a mitochondrial dye (5,5′,6,6′-tetrachloro-1,1,3,3′- tetraethylbenzimidazolylcarbocyanine chloride). The BDTM MitoScreen Kit (BD Biosciences, USA) stains mitochondria in living cells in a membrane potential-dependent fashion. The JC-1 monomer is in equilibrium with J-aggregates that bind to the mitochondrial membrane. The monomer JC-1 fluoresces green (λ em = 527 nm), while the J-aggregates fluoresce red (λ em = 590 nm). Therefore, cells with normal mitochondrial membrane potential fluoresce orange. The depolarization of the mitochondria results in a decrease in the red component and, therefore, green fluorescence. The experiments were in strict accordance with the BD mitochondrial membrane potential detection kit. In summary, after the cells were trypsinized and washed twice with PBS, the cells were labeled with the fluorescent dye JC-1 for 30 min at 37°C. The excess dye was then removed with assay buffer, and the remaining cells were suspended in the buffer solution. The cells were then observed under a fluorescence microscope after 1 h.
Mitoscreen kit
Manufactured by BD
Sourced in United States
The BD™ MitoScreen Kit is a laboratory equipment product designed for the analysis of mitochondrial function. It provides the core function of enabling researchers to assess various parameters of mitochondrial activity and health in a sample.
Automatically generated - may contain errors
Lab products found in correlation
Facscalibur flow cytometer, by BD (4 mentions)
Facscanto 2, by BD (3 mentions)
Mitotracker red, by Thermo Fisher Scientific (3 mentions)
Facsverse flow cytometer, by BD (2 mentions)
Facscalibur, by BD (2 mentions)
Facsdiva software, by BD (2 mentions)
Facsaria, by BD (1 mentions)
Facsaria 3, by BD (1 mentions)
Cflow software, by BD (1 mentions)
Facs calibur flow cytometry machine, by BD (1 mentions)
Cytoflex, by Beckman Coulter (1 mentions)
Accuri c6 cytometer, by BD (1 mentions)
Fcs express 4, by BD (1 mentions)
Facs caliber system, by BD (1 mentions)
Facsaria 3 sorter, by BD (1 mentions)
Facscanto 2 flow cytometer, by BD (1 mentions)
Facs analysis, by BD (1 mentions)
Annexin 5 fitc propidium iodide detection kit, by Merck Group (1 mentions)
Macs quantify version 2, by Miltenyi Biotec (1 mentions)
Cellquest pro software, by BD (1 mentions)
48 protocols using mitoscreen kit
1
Mitochondrial Membrane Potential Assay
2
Apoptosis Assay with MitoScreen Kit
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Apoptosis assay was performed using BDTM MitoScreen Kit (BD Biosciences, San Jose, CA) according to manufacturer’s protocol. Briefly, 3 x 104 cells were seeded in 6-well plates and treated with root extracts with the concentrations of 20 to 40 μg/mL for three days. Cells were trypsinized and transferred into a sterile tube. The cells were centrifuged at 400 g for 5 min at room temperature. Supernatant was discarded and 500 μL of JC-1 solution was added. Cell was incubated at 37 °C for 15 min. After that, cells were washed twice using 1x assay buffer and centrifuged at 400 g for 5 min. Cells were resuspended with 500 μL of 1x assay buffer and analyzed using BD FACSAria flow cytometer (BD Bioscience, San Jose, CA).
3
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetrathylbenzimidazolcarbocyanine iodide) dye from BDTM MitoScreen kit ((BD Bioscience, San Jose, CA, USA) was used to evaluate the status of mitochondrial membrane potential of the cells. After exposure to Streptomyces sp. MUM256 EA for a designated period, cells were harvested and washed twice before being subjected to 10,000 events acquisition and analysis by using BD FACSVerseTM flow cytometer (BD Bioscience, San Jose, CA, USA).
4
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 analyzed with BDTM MitoScreen Kit (BD Bioscience) according to the manufacturer’s instructions. Cells were stained with JC-1 solution for 30 min and analyzed with an Attune NxT Flow Cytometer using two different filter sets: the one at excitation/emission of 488/530 nm for detecting polarized mitochondria and the other at excitation/emission of 561/585 nm for detecting depolarized mitochondria.
5
Mitochondrial Membrane Potential Analysis of CD4+ T Cells
6
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The depolarization of the mitochondrial membrane potential was measured by using the BD MitoScreen Kit (BD Biosciences). Bone cancer cell lines were seeded in a 6-well plate with a density of 6 × 104 cells/well. The next day, the cells were exposed to three different concentrations of DK1. After 48 h, the cells were collected and centrifuged at 2000× g for 5 min. Around 1 × 106 cells were incubated with 500 μL of JC-1 working solution. The JC-1 working solution was prepared in accordance of 1:100 ratios of JC-1 stock solution and assay buffer. This working solution was incubated at 37 °C for 15 min. Then, the cells were washed twice using the assay buffer, before proceeding to the BD Accuri™ C6 analysis (Becton Dickinson).
7
Assessing Mitochondrial Membrane Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The loss of Δ Ψm was assessed using the BD MitoScreen kit according to manufacturer's instructions. The cells were seeded at a density of 2.7×105 cells in 6-well plates for overnight and then treated with FKC at the concentrations of 40, 60, and 80 μM for 24 and 48 h. Following treatment, the cells were harvested and washed twice with PBS. The cells were then suspended in 500 μL of JC-1 working solution and incubated at 37°C for 30 min. A total of 10,000 events were recorded by flow cytometry and analyzed by the BD CFlow software.
8
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
JC-1 assay was used to detect the depolarization of the mitochondrial membrane potential of the cell after being treated with the selected doses of NLC-Citral for 48 hours using BD MitoScreen Kit (BD Pharmingen, USA). In brief, the cells were seeded in 6 wells plate for 24 hours at a density of 1.8 × 105 cells per well. Then, the cells were treated with various concentrations described in Table 1 . This assay was performed based on the manufacturer’s instruction provided with the kit without any modifications. After 48 hours of incubation, the cells were harvested, collected, and incubated with 500 µL of JC-1 stock solution for 15 minutes. Then, it was preceded for analysis by BD FACS Calibur (Becton Dickinson, USA). The result was analyzed using CellQuest 3.3 software. The experiment was performed in triplicates81 (link).
9
Mitochondrial Membrane Potential Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
JC-1 assay was used to detect the depolarization of the mitochondrial membrane potential of the cell after being treated with the selected doses of DMCH for 48 h. This assay was accomplished using a BD MitoScreen Kit (BD Pharmingen, San Diego, CA, USA). In brief, the cells were seeded in a 6-well plate for 24 h at a density of 2.0 × 105 cells per well. Then, the cells were treated with various concentrations as described in Table 1 . This assay was performed based on the manufacturer’s instructions provided with the kit without any modification. After 48 h of incubation, the cells were harvested, collected, and incubated with 500 μL of JC-1 stock solution for 15 min. Then, analysis was performed using a Novocyctes flow cytometer (ACEA Biosciences Inc., San Diego, CA, USA).
10
Mitochondrial Membrane Potential Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The influence of the treatments with the gallates in the mitochondrial membrane potential was assessed. The parasites were treated with the substances during 72 hours using, as standard concentration, the IC50 previously determined by the MTT assay. The percentage of parasite cells which suffered loss in the mitochondrial membrane potential due to the treatment with the tested compounds was measured by flow cytometry using JC-1 dye (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide, BD MitoScreen kit) according to the manufacturer's instruction [27 (link), 28 (link)]. A FacsCanto I cytometer was used, and the data were recorded and analyzed on the software of the equipment. Pentamidine was used as positive control, at 58.7 μM for 24-hour treatment.
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!