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Mitochondrial membrane potential detection kit

Manufactured by Cayman Chemical
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The Mitochondrial Membrane Potential Detection Kit is a laboratory tool used to measure the electrochemical gradient across the inner mitochondrial membrane, which is a key indicator of mitochondrial function. The kit provides the necessary reagents and protocols to assess this important parameter in cell-based research applications.

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Lab products found in correlation

Fl600 fluorescent plate reader, by Agilent Technologies (1 mentions)

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JC-1 mitochondrial membrane potential detection kit

5 protocols using mitochondrial membrane potential detection kit

1

Mitochondrial Membrane Potential Assay in HBMECs

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The change in mitochondrial membrane potential in the HBMECs was monitored using the mitochondrial membrane potential detection kit (Cayman Chemical Company) according to the manufacturer’s instructions. Briefly, HBMECs cultured in either 24-well plate (1×105 cells per well) or 96-well plate (3×104 cells per well) were treated with Tat followed by treatment with 1× JC-1 reagent diluted in serum-free culture medium for 20 min at 37 °C in 5 % CO2. Thereafter, cells were rinsed once in 1× rinsing buffer provided in the kit. Fluorescence was measured using the FL600 fluorescent plate reader (Bio-Tek Instruments, Winooski, VT) at the excitation wavelengths of 485 and 535 nm. All experiments were repeated at least three times.
Ma R., Yang L., Niu F, & Buch S. (2014). HIV Tat-Mediated Induction of Human Brain Microvascular Endothelial Cell Apoptosis Involves Endoplasmic Reticulum Stress and Mitochondrial Dysfunction. Molecular neurobiology, 53(1), 132-142.
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2

Measuring Mitochondrial Membrane Potential

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Changes in the mitochondrial transmembrane potential (ΔΨm) were measured using a JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbo cyanine iodide) mitochondrial membrane potential detection kit (Cayman Chemical Co., Ann Arbor, MI). Cells were exposed to 1 μM valinomycin for 2 h prior to incubation with the JC-1 reagent as a positive control. Cells were incubated with the mitochondrial membrane potential-sensitive fluorescent dye JC-1 for 20 min at 37°C, and then immediately analyzed with flow cytometry, using the 530 nm (FL-1 channel, green) and 585 nm (FL-2 channel, red) band-pass filters simultaneously. Healthy cells with functional mitochondria and high ΔΨm exhibit red fluorescence while apoptotic or dying cells with collapsed mitochondria or low ΔΨm show green fluorescence.
Song G., Valdez B.C., Li Y., Liu Y., Champlin R.E, & Andersson B.S. (2014). Synergistic cytotoxicity of sorafenib with busulfan and nucleoside analogs in human FLT3-ITD-positive acute myeloid leukemia cells. Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, 20(11), 1687-1695.
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3

Evaluating Mitochondrial Membrane Potential

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A mitochondrial membrane potential detection kit (Cayman Chemical Co., Ann Arbor, MI, USA) was used to determine changes in the mitochondrial membrane potential (Δψm) using the JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide) reagent. J45.01 cells to be analyzed were aliquoted (0.5 ml) into 5 ml tubes. Valinomycin (1 μM) was added to a positive control tube containing untreated cells and incubated at 37 °C, 5% CO2 for 15 min. Diluted (1:10 with cell growth medium, 40 μl) Δψm-sensitive fluorescent dye JC-1 reagent was added to each tube, incubated at 37 °C for 20 min, and immediately analyzed by flow cytometry as described by the manufacturer.
Valdez B.C., Zander A.R., Song G., Murray D., Nieto Y., Li Y., Champlin R.E, & Andersson B.S. (2014). Synergistic cytotoxicity of gemcitabine, clofarabine and edelfosine in lymphoma cell lines. Blood Cancer Journal, 4(1), e171-.
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4

Measuring Mitochondrial Membrane Potential

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Changes in the mitochondrial transmembrane potential (ΔΨm) were measured using a JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbo cyanine iodide) mitochondrial membrane potential detection kit (Cayman Chemical Co., Ann Arbor, MI). Cells were exposed to 1 μM valinomycin for 2 h prior to incubation with the JC-1 reagent as a positive control. Cells were incubated with the mitochondrial membrane potential-sensitive fluorescent dye JC-1 for 20 min at 37°C, and then immediately analyzed with flow cytometry, using the 530 nm (FL-1 channel, green) and 585 nm (FL-2 channel, red) band-pass filters simultaneously. Healthy cells with functional mitochondria and high ΔΨm exhibit red fluorescence while apoptotic or dying cells with collapsed mitochondria or low ΔΨm show green fluorescence.
Ji J., Valdez B.C., Li Y., Liu Y., Teo E.C., Nieto Y., Champlin R.E, & Andersson B.S. (2016). Cladribine, gemcitabine, busulfan and SAHA combination as a potential pre-transplant conditioning regimen for lymphomas: a preclinical study. Experimental hematology, 44(6), 458-465.
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5

Mitochondrial Transmembrane Potential Assay

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Changes in the mitochondrial transmembrane potential (ΔΨm) were measured using a JC-1 (5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbo cyanine iodide) mitochondrial membrane potential detection kit (Cayman Chemical Co., MI). KBM3/Bu2506 and OCI-AML3 cells were exposed to 1 μM valinomycin for 4 h prior to incubation with the JC-1 reagent as positive controls. Cells were incubated with the mitochondrial membrane potential-sensitive fluorescent dye JC-1 for 20 min at 37°C, and then immediately analyzed with a flow cytometer for signal recording using the 530 nm (FL-1 channel, green) and 585 nm (FL-2 channel, red) band-pass filters simultaneously. Healthy cells with functional mitochondria and high ΔΨm exhibit red fluorescence while apoptotic or dying cells with collapsed mitochondria or low ΔΨm show green fluorescence. Each determination was based on the ratio of red to green mean fluorescence intensities measured in arbitrary units from at least 10 000 cells per sample [31 (link)].
Song G., Valdez B.C., Li Y., Dominguez J.R., Corn P., Champlin R.E, & Andersson B.S. (2014). The histone deacetylase inhibitor SAHA sensitizes acute myeloid leukemia cells to a combination of nucleoside analogs and the DNA-alkylating agent busulfan. Leukemia & lymphoma, 55(7), 1625-1634.
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