Assessment of inflammasome formation by flow cytometry

Sester, David P., Zamoshnikova, Alina, Thygesen, Sara J., Vajjhala, Parimala R., Cridland, Simon O., Schroder, Kate and Stacey, Katryn J. (2016). Assessment of inflammasome formation by flow cytometry. In Coligan, John E., Bierer, Barbara, Margulies, David H., Shevach, Ethan M. and Strober, Warren (Ed.), Current protocols in immunology (pp. 14.40.1-14.40.29) Hoboken, NJ United States: John Wiley & Sons. doi:10.1002/cpim.13

Author Sester, David P.
Zamoshnikova, Alina
Thygesen, Sara J.
Vajjhala, Parimala R.
Cridland, Simon O.
Schroder, Kate
Stacey, Katryn J.
Title of chapter Assessment of inflammasome formation by flow cytometry
Title of book Current protocols in immunology
Place of Publication Hoboken, NJ United States
Publisher John Wiley & Sons
Publication Year 2016
Sub-type Chapter in reference work, encyclopaedia, manual or handbook
DOI 10.1002/cpim.13
Open Access Status Not Open Access
Year available 2016
Series Current Protocols in Immunology
ISBN 9780471142737
Editor Coligan, John E.
Bierer, Barbara
Margulies, David H.
Shevach, Ethan M.
Strober, Warren
Chapter number 14.40
Start page 14.40.1
End page 14.40.29
Total pages 29
Total chapters 384
Collection year 2017
Language eng
Abstract/Summary Inflammasomes are large protein complexes formed in response to cellular stresses that are platforms for recruitment and activation of caspase 1. Central to most inflammasome functions is the adapter molecule ASC (apoptosis-associated speck-like protein containing a caspase-recruitment domain) that links the inflammasome initiator protein to the recruited caspases. ASC is normally diffuse within the cell but within minutes of inflammasome activation relocates to a dense speck in the cytosol. The dramatic redistribution of ASC can be monitored by flow cytometry using parameters of fluorescence peak height and width when immunostained or tagged with a fluorescent protein. This can be used to define cells with active inflammasomes within populations of primary macrophages and monocytes, allowing quantification of responses and flow-sorting of responding cells. Protein structural requirements for ASC speck formation and recruitment of caspases to ASC specks can be assessed by expressing components in HEK293 cells. This provides rapid quantification of responding cell number and correlation with the expression level of inflammasome components within single cells.
Keyword ASC
Flow cytometry
Q-Index Code BX
Q-Index Status Provisional Code
Institutional Status UQ

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Created: Fri, 05 Aug 2016, 10:13:16 EST by Mrs Louise Nimwegen on behalf of School of Chemistry & Molecular Biosciences