Inducing apoptosis using chemical treatment and acidic pH, and detecting it using the Annexin V flow cytometric assay
Cell death is important in physiology, and can happen as a result of structural damage, or as a sequence of programmed cellular processes known as apoptosis. Pathogenic alterations in apoptosis occur in a number of diseases, including cancer, viral infections, autoimmune diseases, immunodeficiencies...
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| Veröffentlicht in: | PloS one 2022-06, Vol.17 (6), p.e0270599-e0270599 |
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| description | Cell death is important in physiology, and can happen as a result of structural damage, or as a sequence of programmed cellular processes known as apoptosis. Pathogenic alterations in apoptosis occur in a number of diseases, including cancer, viral infections, autoimmune diseases, immunodeficiencies, and degenerative conditions. Developing accurate and reproducible laboratory methods for inducing and detecting apoptosis is vital for research into these conditions. A number of methods are employed to detect cell death, including DNA fragmentation, the TUNEL assay, and electron microscopy although each has its limitations. Flow cytometry allows for the distinction between live, early apoptotic, late apoptotic and necrotic cells. In this protocol we successfully induce apoptosis using chemical treatment and treatment with low pH in solid tumour cell lines, and have optimized detection using the Annexin V/PI apoptosis assay. |
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Pathogenic alterations in apoptosis occur in a number of diseases, including cancer, viral infections, autoimmune diseases, immunodeficiencies, and degenerative conditions. Developing accurate and reproducible laboratory methods for inducing and detecting apoptosis is vital for research into these conditions. A number of methods are employed to detect cell death, including DNA fragmentation, the TUNEL assay, and electron microscopy although each has its limitations. Flow cytometry allows for the distinction between live, early apoptotic, late apoptotic and necrotic cells. In this protocol we successfully induce apoptosis using chemical treatment and treatment with low pH in solid tumour cell lines, and have optimized detection using the Annexin V/PI apoptosis assay.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0270599</identifier><identifier>PMID: 35767593</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Analysis ; Annexin A5 - metabolism ; Annexin V ; Annexins ; Apoptosis ; Assaying ; Autoimmune diseases ; Biology and Life Sciences ; Cancer ; Cell culture ; Cell death ; Cellular structure ; Chemical treatment ; Deoxyribonucleic acid ; DNA ; DNA fragmentation ; Electron microscopy ; Flow cytometry ; Flow Cytometry - methods ; Humans ; Hydrogen-Ion Concentration ; Lab Protocol ; Laboratories ; Laboratory methods ; Medical prognosis ; Medicine and Health Sciences ; Morphology ; Necrosis ; pH effects ; Plasma ; Research and Analysis Methods ; Solid tumors ; Structural damage ; Tumor cell lines</subject><ispartof>PloS one, 2022-06, Vol.17 (6), p.e0270599-e0270599</ispartof><rights>COPYRIGHT 2022 Public Library of Science</rights><rights>2022 Worsley et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 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| subjects | Analysis Annexin A5 - metabolism Annexin V Annexins Apoptosis Assaying Autoimmune diseases Biology and Life Sciences Cancer Cell culture Cell death Cellular structure Chemical treatment Deoxyribonucleic acid DNA DNA fragmentation Electron microscopy Flow cytometry Flow Cytometry - methods Humans Hydrogen-Ion Concentration Lab Protocol Laboratories Laboratory methods Medical prognosis Medicine and Health Sciences Morphology Necrosis pH effects Plasma Research and Analysis Methods Solid tumors Structural damage Tumor cell lines |
| title | Inducing apoptosis using chemical treatment and acidic pH, and detecting it using the Annexin V flow cytometric assay |
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