Mitochondrial membrane potential was measured using the MitoProbe™ JC-1 (Molecular Probes, Eugene, OR) assay kit following the protocol provided. Mitochondrial ROS was measured using the MitoSOX™ Red kit (Molecular Probes, Eugene, OR) according to the manufacture’s guide. Five μM antimycin A was added 10 minutes after (SML0737, Sigma, St Louis, MO) or 5 μM of MitoTEMPO was added an hour prior to the start of the assay for positive and negative controls, respectively.
Mitoprobe jc 1
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The MitoProbe JC-1 is a fluorescent probe used to detect changes in mitochondrial membrane potential. It can be used to assess mitochondrial function in various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Mitotracker red cmxros, by Thermo Fisher Scientific (3 mentions)
Facsverse, by BD (3 mentions)
Pgc 1α, by Abcam (2 mentions)
Bcl 2, by Abcam (2 mentions)
Gapdh, by Abcam (2 mentions)
Bcl2 associated x (bax), by Abcam (2 mentions)
Tim23, by Abcam (2 mentions)
Mttfa, by Abcam (2 mentions)
Caspase 3 activity assay kit, by Abcam (2 mentions)
Phalloidin tritc, by Merck Group (2 mentions)
Annexin 5 fitc, by Beyotime (2 mentions)
Mitosox red mitochondrial superoxide indicator, by Thermo Fisher Scientific (2 mentions)
Lsm 900, by Zeiss (2 mentions)
Mitosox red kit, by Thermo Fisher Scientific (1 mentions)
Sml0737, by Merck Group (1 mentions)
Cleaved caspase 3, by Abcam (1 mentions)
Parkin, by Abcam (1 mentions)
Cox 4, by Abcam (1 mentions)
Luciferase based assay kit, by Promega (1 mentions)
Caspase 1, by Abcam (1 mentions)
22 protocols using mitoprobe jc 1
1
Mitochondrial Membrane Potential and ROS
2
Measuring Mitochondrial Membrane Potential
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MMP was determined by the MitoProbe™ JC-1 (Molecular Probes, Eugene, OR, USA) according to the manufacturer’s instructions. At the end of the 30 h incubation periods, all cochlear explants were washed with PBS and incubated with JC-1 (2 µM) for 50 min in 5% CO2 at 37 °C in the dark. To examine the sensitivity of JC-1, 50 mM of carbonyl cyanide 3-chlorophenylhydrazone (CCCP) were added to the cochlear explants that were not treated with any drugs before treatment of JC-1 and incubated for 5 min at 37 °C. We quantified red over green JC-1 fluorescence ratios by splitting the images according to color channels and measured the average grayscale using Image J software (http://imagej.nih.gov/ij/ ).
3
Mitochondrial Proteome Profiling and Function
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For extraction of mitochondrial and cytosolic proteins, a commercially available kit from Sigma-Aldrich (#MITOISO2, Saint Louis, MO, USA) was used. For staining of mitochondria, MitoProbe™ JC-1 (5,5ʹ,6,6ʹ-tetrachloro-1,1ʹ,3,3ʹ-tetraethyl-imidacarbocyanine iodide) (#M34152, Molecular Probes, Invitrogen, CA) was used. A luciferase-based assay kit was purchased from Promega Corp. (#G7570, Madison, WI). Parkin (#ab77924), TOM20 (#ab186735), TIM23 (#ab230253), PGC-1α (#ab191838), mt-TFA (#ab131607), LC3 (#ab51520), P62 (#ab56416), P62 phospho S349 (#ab211324), COX IV (#ab202554), NLRP3 (#ab214185), ASC (#ab175449), caspase-1 (#ab179515), cleaved caspase-3 (#ab49822), IL-1β (#ab9722), KIM-1 (#ab47635), Bax (#ab32503), Bcl-2 (#ab182858) and GAPDH (#ab181602) antibodies were purchased from Abcam (Cambridge, UK). The CellTiter-Glo assay and terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) staining kit were obtained from Keygen Biotech (#KGA7037, Nanjing, China). ELISA kits to assay inflammatory cytokines (tumour necrosis factor-α [TNF-α, #1217202], interleukin-1β [IL-1β, #1210122], and interleukin-6 [IL-6, #1210602]) were obtained from Dakewe Biotech (Shenzhen, Guangdong, China). The immunohistochemical kits were obtained from Beyotime (#P0203, Shanghai, China), and all the other chemicals were purchased from Sigma-Aldrich (Saint Louis, MO, USA).
4
Mitochondrial Dysfunction Evaluation in Lung Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MitoProbe™ JC-1 (5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide) were purchased from Molecular Probes (Invitrogen, CA, USA). The CellTiter-Glo® assay and a terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining kit were supplied from Promega Corp. (Madison, WI, USA). Human lung alveolar epithelial cell lines (A549) were obtained from Guangzhou Cellcook Biotech Co., Ltd. (Guangzhou, China). Caspase 3 activity assay kit was obtained from Biovision (San Francisco, USA). Immunohistochemical kits were provided by EnVision™ (Dako, Copenhagen, Denmark). LPO kit was obtained from Cayman Chemical Co. (Ann Arbor, Michigan, USA). SOD and MDA kits were obtained from Jiancheng (Nanjing, China). LPS (E. coli serotype O111:B4), Mdivi-1, and other chemicals were purchased from Sigma-Aldrich (Saint Louis, MO, USA).
5
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Changes in membrane potential were measured using the Mitoprobe JC-1 from Molecular Probes® (Eugene OR, USA). The mitochondrial membrane disrupter CCP at 50 µM was used as positive control. The fluorescence intensity of JC-1-stained infected red cells was calculated through flow cytometry using the FloMax software.
6
Mitochondrial Dysfunction and Apoptosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PD was supplied by Neptunus Co. (Shenzhen, Guangdong, China), and MitoProbe™ JC-1 (5,5′,6,6′-Tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide) was purchased from Molecular Probes (Invitrogen, CA). Mitochondriatargeted Keima plasmid (mt-Keima-COX8) was purchased from Public Protein/Plasmid Library (Nanjing, China). siRNA targeting Parkin and Atg7 were purchased from Santa Cruz Biotechnology. The terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) staining kit was supplied by Promega Corp. (Madison, WI). A mitochondrial/ cytosolic protein extraction kit was purchased from BestBio Co. (Beijing, China). Antibodies against Parkin, COX4I1, TOM20, TIM23, PGC-1α, mt-TFA, LC3I/LC3II, Bax, Bcl-2, cytochrome c and GAPDH were obtained from Abcam (Cambridge, UK). The Caspase-3 activity assay kit was obtained from Biovision (San Francisco, USA). Immunohistochemical kits were provided by EnVision™ (Dako, Copenhagen, Denmark). The fluorescein isothiocyanate (FITC) Annexin V apoptosis kit was obtained from BD Biosciences (San Jose, CA). Human lung epithelial cell lines (Beas-2B) were obtained from Guangzhou Cellcook Biotech Co., Ltd (Guangzhou, China). All other chemicals were acquired from Sigma-Aldrich (Saint Louis, MO, USA).
7
Mitochondrial Characterization of Epithelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The epithelial single-cell suspension was incubated with 100 nM MitoTracker Green, 5 μM MitoSOX Red, and 2 μM MitoProbe JC-1 (Invitrogen) for 30 min at 37°C to measure mitochondrial mass, mROS, and membrane potential, respectively. Dead cells were detected with LIVE/DEAD Fixable Violet Dead Cell Stain kit (Invitrogen), followed by incubation with mouse Fc blocking reagent (Miltenyi Biotec) and APC/Cy7-EpCAM antibody (BioLegend). Mean fluorescence intensity in EpCAM+ iECs was quantified by flow cytometry. mtDNA copies were measured by quantitative real-time PCR (qPCR) comparing copies of mitochondrially encoded mt-Nd1 to those of chromosomally encoded β-globin.
8
Apoptosis and Mitochondrial Damage in T Cells
9
Mitochondrial Membrane Potential Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The qualitative and quantitative detection of changes in mitochondrial membrane potential during cell death was performed by MitoProbe JC‐1 (Invitrogen). Breast cancer cells (15 000 cells per well) were pretreated with 3‐MA (5 mm ) for 24 h, then loaded on 100 µL JC‐1 staining working solution, and incubated at 37 °C for 20 min in the cell culture incubator. Washing twice with JC‐1 staining buffer (1×), the absorbance was measured by a multifunctional microplate reader (PerkinElmer) with excitation light at 490 nm and the emission light at 530 nm for JC‐1 monomer detection, while the excitation light at 525 nm and the emission light at 590 nm for JC‐1 polymer detection. Red fluorescence‐labeled mitochondria with high potentials and green fluorescence‐labeled mitochondria with low potentials, which is an indicator fluorescence‐labeled of apoptosis. The red‐green ratio was calculated as indicated.
10
Mitochondrial Membrane Potential Measurement
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mitochondrial membrane potential was estimated using the cationic fluorescent dye MitoProbe™ JC-1 (Invitrogen, Eugene, OR, USA) according to the manufacturer's instructions. At the end of the 3 or 5-day incubation period, all cochlear explants were washed with PBS and incubated with 2 μM JC-1 for 50 min in the dark. To confirm the sensitivity of JC-1, the cochlear explants were not treated with any drugs, 50 μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was added before treatment with JC-1, and the samples were incubated. If the cells have a normal range of mitochondrial membrane potentials, fluorescence emission shift from green to red.
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!