Abstract
Mitochondria play a central role in programmed cell death through the release of cytochrome c and other proapoptotic factors. Fluorescence microscopy is used to visualize cytochrome c translocation and loss of mitochondrial membrane potential. Flow cytometry can also be used to measure mitochondrial membrane potential. Cytochrome c content in cytosol and mitochondria can be determined by immunoblotting after subcellular fractionation or selective permeabilization with digitonin. Isolated mitochondria can be used to study the mechanism of cytochrome c release. This article summarizes some of the more widely used methods to assess mitochondrial alterations in apoptosis.
MeSH terms
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Apoptosis*
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BH3 Interacting Domain Death Agonist Protein
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Carrier Proteins / metabolism
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Cell Death
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Cell-Free System
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Cytochrome c Group / analysis
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Cytosol / metabolism
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Dose-Response Relationship, Drug
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Flow Cytometry
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Green Fluorescent Proteins
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HeLa Cells
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Humans
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Immunoblotting
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Jurkat Cells
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Luminescent Proteins / metabolism
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Membrane Potentials
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Microscopy, Fluorescence
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Mitochondria / metabolism*
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Mitochondria / pathology*
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Potassium / pharmacology
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Protein Transport
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Proto-Oncogene Proteins / metabolism
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Proto-Oncogene Proteins c-bcl-2*
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Recombinant Fusion Proteins / metabolism
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Subcellular Fractions / metabolism
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bcl-2-Associated X Protein
Substances
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BH3 Interacting Domain Death Agonist Protein
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BID protein, human
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Carrier Proteins
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Cytochrome c Group
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Luminescent Proteins
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Proto-Oncogene Proteins
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Proto-Oncogene Proteins c-bcl-2
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Recombinant Fusion Proteins
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bcl-2-Associated X Protein
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Green Fluorescent Proteins
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Potassium