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A real-time fluorometric method for the simultaneous detection of cell death type and rate

Sasker Grootjans (UGent) , Behrouz Hassannia (UGent) , Iris Delrue (UGent) , Vera Goossens (UGent) , Bartosz Wiernicki (UGent) , Yves Dondelinger (UGent) , Mathieu Bertrand (UGent) , Dmitri Krysko (UGent) , Marnik Vuylsteke (UGent) , Peter Vandenabeele (UGent) , et al.
(2016) NATURE PROTOCOLS. 11(8). p.1444-1454
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Abstract
Several cell death assays have been developed based on a single biochemical parameter such as caspase activation or plasma membrane permeabilization. Our fluorescent apoptosis/necrosis (FANAN) assay directly measures cell death and distinguishes between caspase-dependent apoptosis and caspase-independent necrosis of cells grown in any multiwell plate. Cell death is monitored in standard growth medium as an increase in fluorescence intensity of a cell-impermeable dye (SYTOTOX Green) after plasma membrane disintegration, whereas apoptosis is detected through caspase-mediated release of a fluorophore from its quencher (DEVD-amc). The assay determines the normalized percentage of dead cells and caspase activation per condition as an end-point measurement or in real time (automated). The protocol can be applied to screen drugs, proteins or siRNAs for interference with cell death while simultaneously detecting cell death modality switching between apoptosis and necrosis. Initial preparation may take up to 5 d, but the typical hands-on time is similar to 2 h.
Keywords
FACTOR-INDUCED NECROSIS, KINASE-DEPENDENT APOPTOSIS, ANTITUMOR IMMUNITY, NECROPTOSIS, RIPK3, MLKL, CYTOTOXICITY, INFLAMMATION, ACTIVATION, DEPLETION

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Chicago
Grootjans, Sasker, Behrouz Hassannia, Iris Delrue, Vera Goossens, Bartosz Wiernicki, Yves Dondelinger, Mathieu Bertrand, et al. 2016. “A Real-time Fluorometric Method for the Simultaneous Detection of Cell Death Type and Rate.” Nature Protocols 11 (8): 1444–1454.
APA
Grootjans, S., Hassannia, B., Delrue, I., Goossens, V., Wiernicki, B., Dondelinger, Y., Bertrand, M., et al. (2016). A real-time fluorometric method for the simultaneous detection of cell death type and rate. NATURE PROTOCOLS, 11(8), 1444–1454.
Vancouver
1.
Grootjans S, Hassannia B, Delrue I, Goossens V, Wiernicki B, Dondelinger Y, et al. A real-time fluorometric method for the simultaneous detection of cell death type and rate. NATURE PROTOCOLS. 2016;11(8):1444–54.
MLA
Grootjans, Sasker, Behrouz Hassannia, Iris Delrue, et al. “A Real-time Fluorometric Method for the Simultaneous Detection of Cell Death Type and Rate.” NATURE PROTOCOLS 11.8 (2016): 1444–1454. Print.
@article{8044971,
  abstract     = {Several cell death assays have been developed based on a single biochemical parameter such as caspase activation or plasma membrane permeabilization. Our fluorescent apoptosis/necrosis (FANAN) assay directly measures cell death and distinguishes between caspase-dependent apoptosis and caspase-independent necrosis of cells grown in any multiwell plate. Cell death is monitored in standard growth medium as an increase in fluorescence intensity of a cell-impermeable dye (SYTOTOX Green) after plasma membrane disintegration, whereas apoptosis is detected through caspase-mediated release of a fluorophore from its quencher (DEVD-amc). The assay determines the normalized percentage of dead cells and caspase activation per condition as an end-point measurement or in real time (automated). The protocol can be applied to screen drugs, proteins or siRNAs for interference with cell death while simultaneously detecting cell death modality switching between apoptosis and necrosis. Initial preparation may take up to 5 d, but the typical hands-on time is similar to 2 h.},
  author       = {Grootjans, Sasker and Hassannia, Behrouz and Delrue, Iris and Goossens, Vera and Wiernicki, Bartosz and Dondelinger, Yves and Bertrand, Mathieu and Krysko, Dmitri and Vuylsteke, Marnik and Vandenabeele, Peter and Vanden Berghe, Tom},
  issn         = {1754-2189},
  journal      = {NATURE PROTOCOLS},
  language     = {eng},
  number       = {8},
  pages        = {1444--1454},
  title        = {A real-time fluorometric method for the simultaneous detection of cell death type and rate},
  url          = {http://dx.doi.org/10.1038/nprot.2016.085},
  volume       = {11},
  year         = {2016},
}

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