Jc 1 test kit

Manufactured by Beyotime

Sourced in China

The JC-1 test kit is a laboratory product used to measure the mitochondrial membrane potential (MMP) in cells. It contains the fluorescent dye JC-1, which can detect changes in MMP. The kit provides the necessary reagents and protocols to perform this analysis.

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Lsrfortessa flow cytometer, by BD (1 mentions) β actin antibody, by Thermo Fisher Scientific (1 mentions) Nf κb p65 antibody, by Abcam (1 mentions) Mdivi 1, by Bio-Techne (1 mentions) Collagenase type 2, by Merck Group (1 mentions) Albumin fluorescein isothiocyanate conjugate fitc bsa, by Merck Group (1 mentions) Gapdh antibody, by Thermo Fisher Scientific (1 mentions) Mitotracker deep red fm, by Thermo Fisher Scientific (1 mentions) Lipopolysaccharide lps, by Merck Group (1 mentions) Fluorescence microscope, by Olympus (1 mentions) Calcein am, by Beyotime (1 mentions) Cocl2, by Beyotime (1 mentions)

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JC-1 kit (165 citations)

5 protocols using jc 1 test kit

Mitochondrial Membrane Potential Assay

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The mitochondrial membrane potential was measured using the JC-1 test kit (C2003S, Beyotime, China) following the manufacturer’s instructions. NPCs were stained with a JC-1 staining solution at 37 °C for 20 min after exposure to different stimulations for 24 h. A fluorescence microscope (Olympus, Japan) was used to capture fluorescent images. The ratio of red to green fluorescence was calculated as an indicator of alterations in the potential of the mitochondrial membrane.
Following the manufacturer’s instructions, the JC-1 test kit (C2003S, Beyotime, China) was used to determine the mitochondrial membrane potential. After being exposed to various stimuli for 24 h, NPCs were stained with a JC-1 staining solution at 37 °C for 20 min. Fluorescent pictures were taken using an Olympus fluorescence microscope (Japan). The ratio of red to green fluorescence was estimated as an indicator of alterations in the mitochondrial membrane’s potential.

Xiang Z., Zhang P., Jia C., Xu R., Cao D., Xu Z., Lu T., Liu J., Wang X., Qiu C., Fu W., Li W., Cheng L., Yang Q., Feng S., Wang L., Zhao Y, & Liu X. (2024). Piezo1 channel exaggerates ferroptosis of nucleus pulposus cells by mediating mechanical stress-induced iron influx. Bone Research, 12, 20.

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Measuring Mitochondrial Membrane Potential

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MMP was determined using the JC-1 test kit (Beyotime Biotech, Shanghai, China) according to the manufacturer’s instructions. Briefly, IPEC-J2 cells were treated using the experimental design described previously. The cells were then incubated with 1 mL of the JC-1 working fluid for 30 min at 37 °C in a cell incubator. The supernatant was removed after incubation. The cells were washed twice with JC-1 buffer and then analyzed using a BD LSRFortessa flow cytometer (BD Company, Franklin Lake, NJ, USA).

Ren H., Li K., Min Y., Qiu B., Huang X., Luo J., Qi L., Kang M., Xia P., Qiao H., Chen J., Cui Y., Gan L., Wang P, & Wang J. (2023). Rehmannia glutinosa Polysaccharides: Optimization of the Decolorization Process and Antioxidant and Anti-Inflammatory Effects in LPS-Stimulated Porcine Intestinal Epithelial Cells. Antioxidants, 12(4), 914.

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Mdivi-1 Mitigates LPS-Induced Cardiomyocyte Injury

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Mdivi-1 was obtained from Tocris bioscience (American). Lipopolysaccharide (LPS) were purchased from Sigma (St. Louis, MO, United States). H9C2 cell line was purchased from Biowit Technologies (Shenzhen, China). DMEM were from Gibco/Invitrogen. ATP content and JC-1 test kit were from Beyotime. Mito-Tracker™ Deep Red FM was obtained from Invitrogen. The kits of measurement of TNFα, IL-6, D-lactate, ROS, MDA, and SOD were from Nanjing Jiancheng Co., Ltd. ROS Detection Kit of cells were purchased from Abcam (Cambridge, MA, United States). Albumin-fluorescein isothiocyanate conjugate (FITC-BSA) and collagenase type Ⅱ were purchased from Sigma (St. Louis, MO, United States). NF-κ B p65 antibody (Abcam, 1:1000), NF-κ B p65 phosphorylation antibody (Abcam, 1:1000), GAPDH antibody (Thermo, 1:1000), β-actin antibody (Thermo, 1:7000). Tunel was purchased from Roche.

Zhu Y., Kuang L., Wu Y., Deng H., She H., Zhou Y., Zhang J., Liu L, & Li T. (2021). Protective Effects of Inhibition of Mitochondrial Fission on Organ Function After Sepsis. Frontiers in Pharmacology, 12, 712489.

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Visualizing Mitochondrial Dynamics in IPEC-J2 Cells

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MPTP was visualized by fluorescence microscopy after staining with a combination of calcein AM, CoCl₂ and/or quenching reagents (Beyotime Biological Technology, Shanghai, China). The cell staining method was the same as previously described. Green calcein AM fluorescence was observed at a 505 nm emission wavelength. The opening of the MPTP in IPEC-J2 cells was judged by the change in fluorescence intensity, and strong green fluorescence indicated that the MPTP was normal.
MMP was visualized by fluorescence microscopy after staining with a JC-1 test kit (Beyotime Biological Technology, Shanghai, China). The cell staining method was the same as previously described [28 (link)]. Red aggregated JC-1 was observed at a 580 nm emission wavelength, and green monomeric JC-1 was observed at a 525 nm emission wavelength. The levels of MMP in IPEC-J2 cells were judged by fluorescence intensity; strong red fluorescence indicated that the MMP was normal, and strong green fluorescence indicated that the MMP was destroyed.

Zhang C., Wang Y., Zhang X., Zhang K., Chen F., Fan J., Wang X, & Yang X. (2024). Maintaining the Mitochondrial Quality Control System Was a Key Event of Tanshinone IIA against Deoxynivalenol-Induced Intestinal Toxicity. Antioxidants, 13(1), 121.

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Mitochondrial Membrane Potential Assay

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Operations were carried out according to the JC-1 test kit purchased from Beyotime Biotechnology (Shanghai, China). Red fluorescence was observed at 540 nm excitation wavelength and 580 nm emission wavelength by laser scanning confocal microscope, and the red or strong red fluorescence indicated the MMP was normal. The green fluorescence was observed at 488 nm excitation wavelength and 525 nm emission wavelength, and the green indicated MMP was destroyed; RFU was measured at the same excitation and emission wavelength by a fluorescence enzyme labeling instrument. The ratio of red RFU to green RFU was used to reflect the change of MMP. If MMP was damaged, the ratio reduced.

Liu W., Huang L., Liu X., Zhu L., Gu Y., Tian W., Zhang L., Deng S, & Yu T. (2022). Urocortin I Protects against Myocardial Ischemia/Reperfusion Injury by Sustaining Respiratory Function and Cardiolipin Content via Mitochondrial ATP-Sensitive Potassium Channel Opening. Oxidative Medicine and Cellular Longevity, 2022, 7929784.

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