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Dimethyl sulfoxide

二甲基亚砜

公司名称: Sigma-Aldrich
产品编号: 41640
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Detection of ASC Oligomerization by Western Blotting
Author:
Date:
2017-05-20
[Abstract]  The apoptosis-associated speck-like protein with a caspase-recruitment domain (ASC) adaptor protein bridges inflammasome sensors and caspase-1. Upon inflammasome activation, ASC nucleates in a prion-like manner into a large and single platform responsible for the recruitment and the activation of caspase-1. Active caspase-1 will in turn promote the proteolytic maturation of the pro-inflammatory cytokine IL-1β. ASC oligomerization is direct evidence for inflammasome activation and its detection allows a read-out independent of caspase-1 and IL-1β. This protocol describes how to detect the ... [摘要]  具有半胱天冬酶募集区(ASC)衔接蛋白的凋亡相关斑点样蛋白桥联炎症体传感器和半胱天冬酶-1。在炎性体活化后,ASC以类似朊病毒的方式成核,成为负责募集和激活半胱天冬酶-1的大而单一的平台。活性胱天蛋白酶-1将反过来促进促炎细胞因子IL-1β的蛋白水解成熟。 ASC寡聚化是炎性体激活的直接证据,其检测允许读取与caspase-1和IL-1β无关。该方案描述了如何通过蛋白质印迹检测ASC的寡聚化。

背景 Inflammasomes是大量的多蛋白平台,其感测各种微生物,内源和环境胁迫因子,导致促炎IL-1细胞因子家族的成熟(Martinon等人,2002; Sharma和Kanneganti, 2016)。激活后,炎性细胞传感器通过pyrin结构域(PYD)-PYD同型相互作用募集衔接蛋白ASC。 ASC通过胱天蛋白酶激活和募集域(CARD)-CARD相互作用又结合半胱天冬酶-1,并有利于caspase-1的自我蛋白水解切割,导致IL-1β和IL-18的成熟(Hoss等人。,2016)。 Inflammasome激活引发ASC二聚体的超分子寡聚化成称为“ASC-specks”或“pyroptosome”(Fernandes-Alnemri等人,2007)的大交织原纤维。 ASC-speck / ...

Luminol-based Assay for Detection of Immunity Elicitor-induced Hydrogen Peroxide Production in Arabidopsis thaliana Leaves
Author:
Date:
2015-12-20
[Abstract]  In Arabidopsis thaliana, one of the very early immune-related responses induced after elicitor perception is the oxidative burst, i.e., reactive oxygen species (ROS) generation including superoxide anion and hydrogen peroxide (H2O2). ROS production plays different roles in a wide range of biotic and abiotic stress responses, including the closure of stomata and the regulation of cell expansion. In particular, elicitor-induced H2O2 is produced mainly by the membrane localized NAD(P)H oxidases RESPIRATORY BURST OXIDASE HOMOLOGUE D ... [摘要]  在拟南芥中,在诱导剂感知之后诱导的非常早期的免疫相关反应之一是氧化爆发,即包括超氧阴离子和过氧化氢的活性氧(ROS)产生(H 2 O 2 O 2)。 ROS产生在宽范围的生物和非生物应激反应中起不同的作用,包括气孔的闭合和细胞扩增的调节。特别地,诱导剂诱导的H 2 O 2 O 2主要由膜定位的NAD(P)H氧化酶呼吸道刺激氧化酶同源物D和F产生。在该方案中,我们描述了简单和可再现的基于鲁米诺/过氧化物酶的测定,以检测和评价在拟南芥叶盘中产生的H 2 O 2亚型的免疫相关累积(OGs),鞭毛蛋白(flg22)或延伸因子 - 热不稳定的(EF-Tu-elf18)处理。该方法基于检测在H 2 O 2 O 2存在下在辣根过氧化物酶(HRP) - 催化的鲁米诺分子氧化后产生的激发发光氨分子发出的发光, sub>。发光的水平以及持续时间与由引发的叶盘产生的H 2 O 2 O 2的量成比例。

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