Log In Sign up
The largest database of trusted experimental protocols

Jc 1 dye

Manufactured by BD
Visit Supplier
Sourced in United States

JC-1 dye is a fluorescent probe used for detecting changes in mitochondrial membrane potential. It exhibits potential-dependent accumulation in mitochondria, indicated by a fluorescence emission shift from green to red as the membrane potential increases.

Automatically generated - may contain errors

Lab products found in correlation

Lsm 710 confocal microscope, by Zeiss (3 mentions) Accuri c6 flow cytometer, by BD (3 mentions) Agarose, by Merck Group (2 mentions) Ethidium bromide, by Merck Group (2 mentions) Acridine orange, by Merck Group (2 mentions) Mitotracker red, by BD (2 mentions) Propidium iodide, by Merck Group (2 mentions) Fluo 4 am, by Thermo Fisher Scientific (2 mentions) H2dcfda, by Thermo Fisher Scientific (2 mentions) Matrix metallopeptidase 9 mmp 9, by Santa Cruz Biotechnology (1 mentions) Anti rabbit igg fluorescein isothiocyanate fitc, by Santa Cruz Biotechnology (1 mentions) Cell culture reagents, by Thermo Fisher Scientific (1 mentions) Hepes, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Santa Cruz Biotechnology (1 mentions) Jc 1 dye, by Thermo Fisher Scientific (1 mentions) Tryple, by Thermo Fisher Scientific (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Zen 2011, by Zeiss (1 mentions) Accuri c5, by BD (1 mentions) Accuri c6, by BD (1 mentions)

11 protocols using jc 1 dye

1

Measuring Mitochondrial Membrane Potential

Check if the same lab product or an alternative is used in the 5 most similar protocols
The effect of complex 2 on the mitochondrial membrane potential (ΔΨm) was measured using JC-1 dye (BD Biosciences) by flow cytometry according to the manufacturer's instructions. Briefly, 3.0×105 S180 cells were treated with complex 2 (40 µM and 60 µM) for 24 h. After incubation, the cells were harvested and washed with PBS. The cells were then incubated with JC-1 dye (BD Biosciences) for 15 min at 37°C in the dark. The stained cells were washed, resuspended in assay buffer, and immediately analyzed by flow cytometry. The data were analyzed using the Cell Quest software.
Lima A.P., Pereira F.C., Almeida M.A., Mello F.M., Pires W.C., Pinto T.M., Delella F.K., Felisbino S.L., Moreno V., Batista A.A, & de Paula Silveira-Lacerda E. (2014). Cytoxicity and Apoptotic Mechanism of Ruthenium(II) Amino Acid Complexes in Sarcoma-180 Tumor Cells. PLoS ONE, 9(10), e105865.
+ Open protocol
+ Expand
2

Apoptosis and Inflammatory Signaling Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
EA, TBA, DTNB, HEPES were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies against BCL2, Bax, p53, p21, Matrix metallopeptidase-9 (MMP-9), and anti-rabbit IgG fluorescein isothiocyanate (FITC) and Rhodamine 1,2,3 conjugated secondary antibodies, and 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Santa Cruz (Santa Cruz, CA, USA). 2′,7′-Dichlorofluoroscein diacetate (H2DCF-DA) was purchased from Calbiochem of Merck-Millipore (Billerica, MA, United States). Antibody against p-NF-κB, p-STAT3, cyclooxygenase-2 (COX-2) and p-Akt were purchased from Cell Signaling (Beverly, MA, USA). Antibody to detect the level of active caspase 3, annexin-FITC-PI, and JC1 dye were purchased from Becton Dickinson-Biosciences (San Jose, CA, USA). All the cell culture reagents were purchased from Gibco (Waltham, MA, US) and all other reagents used for this study were of highest quality grade.
Das U., Biswas S., Chattopadhyay S., Chakraborty A., Dey Sharma R., Banerji A, & Dey S. (2017). Radiosensitizing effect of ellagic acid on growth of Hepatocellular carcinoma cells: an in vitro study. Scientific Reports, 7, 14043.
+ Open protocol
+ Expand
3

Evaluating Mitochondrial Function in Neurons

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
To measure mitochondrial size, Prox1::eGFP and DsRed2-mito were co-expressed in DG-like neurons via lentiviral infection. Neurons were fixed in 4% paraformaldehyde and then permeabilized with 0.1% Triton-X100 in TBS. Cells were then blocked in TBS containing 3% donkey serum for 1 h, followed by incubation with DAPI for 15 min. Fluorescence images were acquired using a high-resolution LSM 710 confocal microscope (Carl Zeiss) and were processed with ZEN 2011 software (Carl Zeiss) and Adobe Photoshop CS5 software (Adobe). The size of the mitochondria (DsRed2-mito puncta) was analysed using the Particle Analysis tool in ImageJ software (National Institutes of Health).
For MMP, neurons were incubated with JC-1 dye (Molecular Probes) at 37 °C for 15–30 min29 (link). The cells were dissociated into single cells using TrypLE (Invitrogen), washed three times and then resuspended in 1 ml warm PBS. Green and red fluorescence of JC-1 dye was quantitated using BD FACSCanto II flow cytometer (Becton, Dickinson). Histogram plots of green and red fluorescence were created to determine the red/green intensity ratio using FlowJo 10 software (TreeStar).
Mertens J., Wang Q.W., Kim Y., Yu D.X., Pham S., Yang B., Zheng Y., Diffenderfer K.E., Zhang J., Soltani S., Eames T., Schafer S.T., Boyer L., Marchetto M.C., Nurnberger J.I., Calabrese J.R., Oedegaard K.J., McCarthy M.J., Zandi P.P., Alda M., Nievergelt C.M., Mi S., Brennand K.J., Kelsoe J.R., Gage F.H, & Yao J. (2015). Differential responses to lithium in hyperexcitable neurons from patients with bipolar disorder. Nature, 527(7576), 95-99.
+ Open protocol
+ Expand
4

Naringenin Modulates Cellular Redox, Calcium, and Mitochondrial Dynamics

Check if the same lab product or an alternative is used in the 5 most similar protocols
ROS generation was determined using the fluorogenic probe 2′,7′-dichlorodihydrofluorescein (H2DCFDA; Thermo Fisher; Waltham, MA, USA). The mitochondrial membrane potential (MMP) was determined using the cationic JC-1 dye (BD Biosciences). The Ca2+ concentration was measured using the Ca2+-sensitive fluorescent probe Fluo-4 AM (Thermo Fisher). Cells (5 × 105) were plated in six-well plates, grown to confluence, and treated with naringenin at various concentrations (100, 250, and 500 μM) for indicated times. After incubation, cells were stained with H2DCFDA (10 μM), Fluo-4 AM (3 μg/mL), and JC-1 (5 μg/mL) to determine ROS production, Ca2+ levels, and MMP, respectively. NAC and DPI were used as ROS inhibitors, and BAPTA-AM was used to control the intracellular Ca2+ level. Cells were determined using the BD Accuri C5 flow cytometer and BD Accuri C6 software (version 1.0.264.21, BD Biosciences).
Lee C.W., Huang C.C., Chi M.C., Lee K.H., Peng K.T., Fang M.L., Chiang Y.C, & Liu J.F. (2022). Naringenin Induces ROS-Mediated ER Stress, Autophagy, and Apoptosis in Human Osteosarcoma Cell Lines. Molecules, 27(2), 373.
+ Open protocol
+ Expand
5

Evaluating LEAS-induced Mitochondrial Dysfunction

Check if the same lab product or an alternative is used in the 5 most similar protocols
The effect of LEAS on the ΔΨm of D-17 cells was evaluated by flow cytometry using JC-1 dye (BD Biosciences, San Jose, CA, USA) that allows differentiating healthy cells (red fluorescence) from those with mitochondrial damage (green fluorescence). For this, D-17 cells (4 × 105/mL) were cultured in 24-well plates and treated with LEAS IC50 or vehicle and stained with JC-1 dye for 15 min at 37 °C in the dark, according to the manufacturer’s instructions. The cells were subsequently washed twice with assay buffer, and their fluorescence was measured in a BD AccuriTM C6 flow cytometer (BD Biosciences, San Jose, CA, USA). Data were analyzed using the FlowJo Software v. 10.4.2 (TreeStar Inc., Ashland, OR, USA).
Padilla-Arellanes S., Salgado-Garciglia R., Báez-Magaña M., Ochoa-Zarzosa A, & López-Meza J.E. (2021). Cytotoxicity of a Lipid-Rich Extract from Native Mexican Avocado Seed (Persea americana var. drymifolia) on Canine Osteosarcoma D-17 Cells and Synergistic Activity with Cytostatic Drugs. Molecules, 26(14), 4178.
+ Open protocol
+ Expand
6

Mitochondrial Membrane Potential Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
The changes in mitochondrial membrane potential were monitored using the JC-1 dye (BD Biosciences) in a BD Accuri™ C6 flow cytometer (BD Biosciences) [15 (link)]. The K562 cells (1 × 105 cells/well) were cultured in 96-well plates and treated with γ-thionin IC50 or vehicle for 6, 12, and 24 h. The cells were treated according to the manufacturer’s instructions. The fluorescence was measured in a BD Accuri ™ C6 flow cytometer (BD Biosciences). The data were analyzed using FlowJo software version 10.4 (TreeStar, Inc., San Carlos, CA, USA). Actinomycin D (Sigma-Aldrich, 80 μg/mL) was used as a positive control.
Flores-Alvarez L.J., Jiménez-Alcántar P., Ochoa-Zarzosa A, & López-Meza J.E. (2023). The Antimicrobial Peptide γ-Thionin from Habanero Chile (Capsicum chinense) Induces Caspase-Independent Apoptosis on Human K562 Chronic Myeloid Leukemia Cells and Regulates Epigenetic Marks. Molecules, 28(9), 3661.
+ Open protocol
+ Expand
7

Assessment of Manikya Bhasma's Anticancer Potential

Check if the same lab product or an alternative is used in the 5 most similar protocols
Manikya Bhasma was obtained from local Baidyanath store in Guwahati city, acridine orange, propidium iodide, ethidium bromide, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), agarose, 2,7-dichlorofluorescein diacetate (DCFH-DA), dulbecco’s modified eagle’s medium were purchased from Sigma Aldrich (St. Louis, MO, USA). RNase A, Proteinase K, DMSO, Foetal Bovine Serum (FBS), Penicillin-Streptomycin (100X) antibiotic solution, Phosphate Buffer Saline (PBS), sodium azide, trypan blue, and trypsin were obtained from HiMedia (Mumbai, India). Ethylenediaminetetraacetic acid (EDTA), ethanol, sodium chloride, was purchased from Merck, Germany. Anti-Cyt-c antibodies, Mitotracker Red, and JC-1 dye were obtained from BD-Biosciences (San Jose, USA). The caspase-9 colorimetric kit was obtained from Invitrogen Corporation (Waltham, USA). All the cell culture plates and dishes were purchased from Corning, Lowell, MA, USA. MDAMB-231, DLD-1, HCT-116, MG-63, HeLa cancer cell lines were procured from National Centre for Cell Sciences, Pune, India. All other reagents and chemicals were of analytical grade purity.
Jha S, & Trivedi V. (2020). Manikya Bhasma is a nanomedicine to affect cancer cell viability through induction of apoptosis. Journal of Ayurveda and Integrative Medicine, 12(2), 302-311.
+ Open protocol
+ Expand
8

Mitochondrial Membrane Potential Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Assays were performed with 8 × 104 cells/well in 24-well plates and cells were synchronized and treated as described above. Cells were collected and stained with JC-1 dye (BD Biosciences) for 15 min at 37°C in the dark according to the manufacturer’s instructions. The cells were washed twice with assay buffer, and the fluorescence was measured in a BD AccuriTM C6 flow cytometer (BD Biosciences) using FlowJo software (TreeStar, Inc.). The JC-1 dye allows the differentiation of healthy cells (red fluorescence) from those with mitochondrial damage (green fluorescence).
Jiménez-Alcántar P., López-Gómez R., López-Meza J.E, & Ochoa-Zarzosa A. (2022). PaDef (Persea americana var. drymifolia), a Plant Antimicrobial Peptide, Triggers Apoptosis, and Induces Global Epigenetic Modifications on Histone 3 in an Acute Lymphoid Leukemia Cell Line. Frontiers in Molecular Biosciences, 9, 801816.
+ Open protocol
+ Expand
9

Mitochondrial Depolarization Quantification

Check if the same lab product or an alternative is used in the 5 most similar protocols
Changes in the mitochondrial membrane potential (ΔΨmt) were determined using flow cytometry, FACS Calibur (BD Biosciences, USA) as described earlier [40 (link)]. Treated cells were washed in PBS, trypsinized and stained with 10 μg/ml JC-1 dye (Sigma, USA). After that, stained cells were incubated at 37 °C for 30 mins in dark. The mitochondrial depolarization was estimated by determining the number of cells that shifted from red to green fluorescence, in a flow cytometry (FACS Calibur, BD Biosciences, USA), after staining with JC-1dye, 10,000 cells per sample were analyzed [41 (link)]. For fluorescence microscopic analysis, the treated cells were also incubated with 2 μg/ml JC-1 dye in PBS for 20 mins in 37 °C as described in [42 (link)]. The cells were then observed under the fluorescence microscope (Invitrogen EVOS FL Auto Cell Imaging System).
Hansda S., Ghosh G, & Ghosh R. (2020). 9-phenyl acridine photosensitizes A375 cells to UVA radiation. Heliyon, 6(9), e04733.
+ Open protocol
+ Expand
10

Apoptosis and Oxidative Stress Pathways

Check if the same lab product or an alternative is used in the 5 most similar protocols
N-acetylcysteine (NAC), propidium iodide, ethidium bromide, acridine orange, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), thiobarbituric acid, 1,19,3,39-tetraethoxypropane, guanidine hydrochloride, agarose, DAPI-containing mounting solution, filipin, and chemiluminescence peroxidase kits were purchased from Sigma (St. Louis, USA). Dimethylsulfoxide (DMSO), Triton X-100, Tween-20, hydrogen peroxide, methanol, β-mercaptoethanol, acrylamide, and bis-acrylamide were obtained from Merck (Boston, USA). Anti-PKCα and anti-cyt c antibodies, Mito-Tracker Red, and JC-1 dye were obtained from BD-Biosciences (San Jose, USA). Anti-5′-nucleotidase antibody was purchased from Cell Signaling Technology (Danvers, USA). Caspase-3 assay kit was from BD Pharmingen (San Jose, USA). Caspase-9 colorometric kit was from Invitrogen Corp. (Waltham, USA). Other reagents and chemicals were of analytical grade purity.
Deka S.J., Mamdi N., Manna D, & Trivedi V. (2016). Alkyl Cinnamates Induce Protein Kinase C Translocation and Anticancer Activity against Breast Cancer Cells through Induction of the Mitochondrial Pathway of Apoptosis. Journal of Breast Cancer, 19(4), 358-371.
+ Open protocol
+ Expand

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

Sign up now

Revolutionizing how scientists
search and build protocols!