炎症体
先天免疫系统
细胞生物学
免疫印迹
劈理(地质)
半胱氨酸蛋白酶1
化学
调节器
炎症
免疫系统
生物
免疫学
生物化学
古生物学
断裂(地质)
基因
出处
期刊:Methods in molecular biology
日期:2023-01-01
卷期号:: 17-26
标识
DOI:10.1007/978-1-0716-3040-2_2
摘要
The NLRP3 inflammasome is a critical component of innate immunity that activates caspase-1 to induce inflammation in response to a wide spectrum of endogenous and exogenous stimuli. NLRP3 inflammasome activation has been shown by assays for the cleavage of caspase-1 and gasdermin D, the maturation of IL-1β and IL-18, and ASC speck formation in innate immune cells such as macrophages and monocytes. Recently, NEK7 has been revealed as an essential regulator for NLRP3 inflammasome activation by forming high-molecular-weight complexes with NLRP3. Blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used to study multi-protein complexes in many experimental systems. Here, we provide a detailed protocol to detect NLRP3 inflammasome activation and NLRP3-NEK7 complex assembly in mouse macrophages by Western blot and BN-PAGE.
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