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Detection of Pyroptosis by Measuring Released Lactate Dehydrogenase Activity

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The Inflammasome

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1040))

Abstract

Pyroptosis is a form of programmed, inflammatory cell death that is dependent on the activation of a cysteine protease caspase-1. Following caspase-1 activation via inflammasomes (including NLRP3, NLRC4, Nlrp1b, and AIM2), cells lose membrane integrity and lyse, releasing lactate dehydrogenase (LDH) that is normally maintained within the cell cytosol. Thus, pyroptosis is distinct from apoptosis, which results in cellular contents being enclosed within membrane blebs during cellular demise. LDH is only released from apoptotic blebs after secondary necrosis occurs. Pyroptosis is distinct from necrosis in that it requires the activity of caspase-1. In this chapter, we describe enzymatic assays for the detection of LDH released by pyroptotic cells using a commercially available kit, as well as a simple and cost-effective method adapted from Decker et al. (J Immunol Methods 115:61–69, 1988).

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© 2013 Springer Science+business media, New York

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Rayamajhi, M., Zhang, Y., Miao, E.A. (2013). Detection of Pyroptosis by Measuring Released Lactate Dehydrogenase Activity. In: De Nardo, C., Latz, E. (eds) The Inflammasome. Methods in Molecular Biology, vol 1040. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-523-1_7

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  • DOI: https://doi.org/10.1007/978-1-62703-523-1_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-522-4

  • Online ISBN: 978-1-62703-523-1

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