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Detection of pyroptosis by measuring released lactate dehydrogenase activity.

Publication ,  Journal Article
Rayamajhi, M; Zhang, Y; Miao, EA
Published in: Methods Mol Biol
2013

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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Published In

Methods Mol Biol

DOI

EISSN

1940-6029

Publication Date

2013

Volume

1040

Start / End Page

85 / 90

Location

United States

Related Subject Headings

  • Macrophages
  • L-Lactate Dehydrogenase
  • Humans
  • Enzyme Activation
  • Developmental Biology
  • Cytotoxicity, Immunologic
  • Bacterial Infections
  • Apoptosis
  • Animals
  • 3404 Medicinal and biomolecular chemistry
 

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Rayamajhi, M., Zhang, Y., & Miao, E. A. (2013). Detection of pyroptosis by measuring released lactate dehydrogenase activity. Methods Mol Biol, 1040, 85–90. https://doi.org/10.1007/978-1-62703-523-1_7
Rayamajhi, Manira, Yue Zhang, and Edward A. Miao. “Detection of pyroptosis by measuring released lactate dehydrogenase activity.Methods Mol Biol 1040 (2013): 85–90. https://doi.org/10.1007/978-1-62703-523-1_7.
Rayamajhi M, Zhang Y, Miao EA. Detection of pyroptosis by measuring released lactate dehydrogenase activity. Methods Mol Biol. 2013;1040:85–90.
Rayamajhi, Manira, et al. “Detection of pyroptosis by measuring released lactate dehydrogenase activity.Methods Mol Biol, vol. 1040, 2013, pp. 85–90. Pubmed, doi:10.1007/978-1-62703-523-1_7.
Rayamajhi M, Zhang Y, Miao EA. Detection of pyroptosis by measuring released lactate dehydrogenase activity. Methods Mol Biol. 2013;1040:85–90.

Published In

Methods Mol Biol

DOI

EISSN

1940-6029

Publication Date

2013

Volume

1040

Start / End Page

85 / 90

Location

United States

Related Subject Headings

  • Macrophages
  • L-Lactate Dehydrogenase
  • Humans
  • Enzyme Activation
  • Developmental Biology
  • Cytotoxicity, Immunologic
  • Bacterial Infections
  • Apoptosis
  • Animals
  • 3404 Medicinal and biomolecular chemistry