| Chapter title |
NLR Proteins
|
|---|---|
| Chapter number | 9 |
| Book title |
NLR Proteins
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3566-6_9 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3564-2, 978-1-4939-3566-6
|
| Authors |
Beilharz, Michael, De Nardo's, Dominic, Latz, Eicke, Franklin, Bernardo S, Michael Beilharz, Dominic De Nardo’s, Eicke Latz, Bernardo S. Franklin, Dominic De Nardo, Nardo, Dominic, Franklin, Bernardo S. |
| Editors |
Francesco Di Virgilio, Pablo Pelegrín |
| Abstract |
Inflammasome assembly results in the formation of a large intracellular protein scaffold driven by the oligomerization of the adaptor protein apoptosis-associated speck-like protein containing a CARD (ASC). Following inflammasome activation, ASC polymerizes to form a large singular structure termed the ASC "speck," which is crucial for recruitment of caspase-1 and its inflammatory activity. Hence, due to the considerably large size of these structures, ASC specks can be easily visualized by microscopy as a simple upstream readout for inflammasome activation. Here, we provide two detailed protocols for imaging ASC specks: by (1) live-cell imaging of monocyte/macrophage cell lines expressing a fluorescently tagged version of ASC and (2) immunofluorescence of endogenous ASC in cell lines and human immune cells. In addition, we outline a protocol for increasing the specificity of ASC antibodies for use in immunofluorescence. |
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