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100% ethanol

Company: PHARMCO-AAPER
Catalog#: 111000200
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Escherichia coli Infection of Drosophila
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Date:
2017-05-05
[Abstract]  Following septic insults, healthy insects, just like vertebrates, mount a complex immune response to contain and destroy pathogens. The failure to efficiently clear bacterial infections in immuno-compromised fly mutants leads to higher mortality rates which provide a powerful indicator for genes with important roles in innate immunity. The following protocol is designed to reproducibly inject a known amount of non-pathogenic E. coli into otherwise sterile flies and to measure the survival of flies after infection. The protocol can be easily adapted to different types of bacteria. [摘要]  在败血症后,健康的昆虫就像脊椎动物一样,会发生复杂的免疫反应,以遏制和破坏病原体。在免疫损害的蝇突变体中未能有效地清除细菌感染导致更高的死亡率,这为在先天免疫中具有重要作用的基因提供了强有力的指标。以下协议被设计为可重复地注射已知量的非致病性E。大肠杆菌进入其他无菌苍蝇,并测量感染后苍蝇的存活率。该方案可以轻松适应不同类型的细菌。

背景 经典的感染实验包括口服感染果蝇(Chakrabarti等人,2016)或用浸在浓缩细菌溶液中的针(Romeo和Lemaitre,2008)。与这些方案不同,我们的实验程序允许我们确定感染部位,并精确控制注射到每只苍蝇中的细菌的剂量。这提供了均匀性和重复性,并且允许我们适应不同实验的细菌负荷(Akbar等人,2011和2016)。

Assay to Measure Interactions between Purified Drp1 and Synthetic Liposomes
Author:
Date:
2017-05-05
[Abstract]  A mitochondrion is a dynamic intracellular organelle that actively divides and fuses to control its size, number and shape in cells. A regulated balance between mitochondrial division and fusion is fundamental to the function, distribution and turnover of mitochondria (Roy et al., 2015). Mitochondrial division is mediated by dynamin-related protein 1 (Drp1), a mechano-chemical GTPase that constricts mitochondrial membranes (Tamura et al., 2011). Mitochondrial membrane lipids such as phosphatidic acid and cardiolipin bind Drp1, and Drp1-phospholipid interactions provide key regulatory mechanisms for mitochondrial division (Montessuit et al., 2010; Bustillo-Zabalbeitia et al., 2014; Macdonald et al., 2014; Stepanyants et al., 2015; ... [摘要]  线粒体是一种动态的细胞内细胞器,主动分裂和融合以控制细胞的大小,数量和形状。线粒体分裂和融合之间的调节平衡是线粒体功能,分布和周转的基础(Roy等,2015)。线粒体分化是由动力蛋白相关蛋白1(Drp1)介导的,其是限制线粒体膜的机械化学GTP酶(Tamura等人,2011)。线粒体膜脂质如磷脂酸和心磷脂结合Drp1,并且Drp1磷脂相互作用提供线粒体分裂的关键调控机制(Montessuit等人,2010; Bustillo-Zabalbeitia等人2014年; Macdonald等人,2014年; Stepanyants等人,2015; Adachi等人,2016)。在这里,我们描述了使用纯化的重组Drp1和具有定义的一组磷脂的合成脂质体定量测量Drp1与脂质的相互作用的生物化学实验。该测定使得可以定义蛋白质 - 脂质相互作用的特异性以及头基和酰基链的作用。

背景 蛋白质和膜脂质的相互作用对于细胞如细胞器分裂中生物膜的重塑至关重要。在线粒体分裂中,Drp1限制线粒体膜并驱动该膜重塑过程。我们最近显示,信号磷脂,磷脂酸与Drp1相互作用,并通过限制线粒体上的组装分裂机制(Adachi等人,2016)产生启动步骤。 Drp1识别磷脂酸的头基和酰基链。为了分析Drp1-磷脂酸结合,我们建立了几种蛋白质 - ...

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