Jc 1 dye

Manufactured by Santa Cruz Biotechnology

Sourced in United States

JC-1 dye is a fluorescent probe used to measure mitochondrial membrane potential in cells. It exhibits potential-dependent accumulation in mitochondria, indicated by a fluorescence emission shift from green (529 nm) to red (590 nm).

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Lab products found in correlation

Annexin v 7aad staining kit, by BioLegend (1 mentions) Valinomycin, by Cayman Chemical (1 mentions)

2 protocols using jc 1 dye

Evaluating Mitochondrial Dynamics and Apoptosis

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Mitochondrial membrane potential (ΔΨm) was detected by a fluorescent JC-1 dye. MCF7 cells transfected with vector control and FBXL20 for 36 h were then either treated with DMSO or 5 μm Camptothecin for 12 h. Cells were washed twice with PBS and resuspended in serum-free DMEM medium containing JC-1 dye (Santacruz, sc-364116) for 30 min at 37 °C and analysed by flow cytometry. The fluorescence emission shift from red to green was determined as a change in mitochondrial membrane potential.
Percentage of apoptotic cells was determined using Annexin-V/7AAD staining kit (Biolegend). MCF7 cells expressing NS or FBXL20 shRNA or PUMA shRNA or BAX shRNA were treated with either DMSO or Doxorubicin 5 μM, 16 h. Cells were collected, washed with PBS/0.1% BSA, and incubated with FITC-conjugated Annexin V and 7AAD. Stained cells were analyzed by flow cytometry.

Manne R.K., Agrawal Y., Malonia S.K., Banday S., Edachery S., Patel A., Kumar A., Shetty P, & Santra M.K. (2021). FBXL20 promotes breast cancer malignancy by inhibiting apoptosis through degradation of PUMA and BAX. The Journal of Biological Chemistry, 297(4), 101253.

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Mitochondrial Membrane Potential Assay

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The mitochondrial membrane potential assay can be used to evaluate cell function and health; thus, cationic carbocyanine dye (JC-1) staining cells were used [35 (link)]. Thus, ConA-stimulated T lymphocytes exposed to IC₅₀ doses of P. alata extract and polyphenols were washed with PBS 0.1 M (pH 7.4). After centrifuging at 300× g for 5 min at room temperature, the cells were labelled with 1 µg of JC-1 dye (Santa Cruz Biotechnology, Dallas, TX, USA), diluted in 100 µL of PBS and incubated at 37 °C for 30 min. The cells were then washed with PBS, centrifuged for 5 min at 300× g and analyzed immediately by flow cytometry. T cells were submitted to valinomycin (Cayman Chemical, Ann Arbor, MI, USA) as a positive assay control at 100 µM. The results are expressed as the percentage of depolarized (green fluorescent) and polarized (orange–red fluorescent) T lymphocyte mitochondrial membrane potential.

Colomeu T.C., de Figueiredo D., de Matos da Silva P., Fernandes L.G, & Zollner R.D. (2022). Antiproliferative and Pro-Oxidant Effect of Polyphenols in Aqueous Leaf Extract of Passiflora alata Curtis on Activated T Lymphocytes from Non-Obese Diabetic (NOD SHILT/J) Mice. Antioxidants, 11(8), 1503.

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