The mitochondrial membrane potential (ΔΨm) was investigated using the MitoCapture™ Apoptosis Detection Kit (BioVision, Mountain View, CA, USA), according to the manufacturer’s protocol. HL-60 cells (5 × 105 cells/mL) were treated with either 10 µM of 11 or 33 µM of cisplatin for 6 h, and then were centrifuged at 500 × g for 5 min and collected. Cell pellets were resuspended in 250 µL of MitoCapture™ solution, incubated for 15 min at 37 °C, and then centrifuged again for 5 min. Pellets were resuspended in 200 µL of incubation buffer and observed via fluorescence microscopy.
Mitocapture apoptosis detection kit
Manufactured by Abcam
Sourced in United States
The MitoCapture™ Apoptosis Detection Kit is a fluorescent assay for the detection of apoptosis. The kit uses a cationic dye that exhibits potential-dependent accumulation in the mitochondria, enabling the visualization of healthy mitochondria. Upon the initiation of apoptosis, the dye is released from the mitochondria into the cytoplasm, allowing for the identification of apoptotic cells.
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Lab products found in correlation
6 protocols using mitocapture apoptosis detection kit
1
Mitochondrial Membrane Potential Assay
2
Mitochondrial Membrane Permeability Assay
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Cells were prepared for FACS as described above and stained using a Mitocapture Apoptosis Detection kit from BioVision with a fluorescent lipophilic cationic reagent that assesses mitochondrial membrane permeability, according to the manufacturer’s recommendations.
3
Mitochondrial Potential Disruption Assay
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The possible disruption of the mitochondrial potential in HCT116 cells by Plumbagin treatment was monitored using the MitoCapture™ Apoptosis Detection kit (BioVision, Milpitas, CA, USA) following the manufacturer’s instructions. This fluorescence-based assay detects the disruption or total loss of the mitochondrial transmembrane potential as one of the earliest intracellular events that occur following stimulation of apoptotic pathways. Cells were imaged immediately using an IX71 fluorescence microscope at the LCCC Microscopy and Imaging Shared Resource. Excitation was induced at either 478 or 507 nm, and emission (Em) was monitored with FITC (Em, 512–535) and rhodamine filters (Em, 540–560 nm).
4
Mitochondrial Membrane Potential Assay
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The mitochondrial membrane potential was determined using a MitoCapture™ Apoptosis Detection Kit (Biovision, Milpitas, CA, USA) according to the manufacturer's suggested protocol. The kit utilizes a cationic dye that detects changes in the mitochondrial transmembrane potential in healthy (red fluorescence) and in apoptotic (green fluorescence) cells. The intensity of cellular fluorescence was analyzed by fluorescence microscopy or by flow cytometry (FACSCalibur, BD Biosciences). The experiments were performed three times independently. All similar results were obtained and a typical data was shown.
5
Evaluating Mitochondrial Membrane Potential
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Changes in mitochondrial membrane potential (MMP) following induction of apoptosis were assessed using a Mito Capture apoptosis detection kit (BioVision) according to the manufacturer's instructions. The status of the MMP was analyzed by using the FL1 channel of a FACScan flow cytometer.
6
Propofol's Effect on rENSCs Proliferation
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The cell proliferation assay was performed using the Quick Cell Proliferation Assay Kit (Biovision, Inc., Milpitas, CA, USA) according to the manufacturer instructions. rENSCs were seeded on 96-well plates at a density of 7 x 10 4 cells per well with 100 µL medium and incubated for 24 h at 37°C in humidified air containing 5% CO 2 . Propofol dissolved in dimethyl sulfoxide (DMSO) (Sigma, St. Louis, MO, USA) was added at concentrations of 1, 10, and 50 µM, while some wells of cells were treated with 0.1% DMSO as controls. To analyze the role of the GABA A receptor in the effect of propofol on cell growth, the GABA A receptor antagonist securinine was added at a final concentration of 0.1 µM. Each sample was tested in triplicate. After culture for 12, 24, 36, and 48 h, 10 µL WST-1 solution was added into each well and cells were incubated for another 3 h. The optical density of each well was measured using an enzyme-linked immunosorbent assay reader at a wavelength of 440 nm.
Apoptosis was detected using the MitoCapture Apoptosis Detection Kit (Biovision) according to the manufacturer instructions. Apoptotic cells were analyzed by flow cytometry with a fluorescein isothiocyanate channel (Ex/Em = 488/530 + 30 nm).
Apoptosis was detected using the MitoCapture Apoptosis Detection Kit (Biovision) according to the manufacturer instructions. Apoptotic cells were analyzed by flow cytometry with a fluorescein isothiocyanate channel (Ex/Em = 488/530 + 30 nm).
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