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Adenosine 5'-diphosphate sodium salt

腺苷5''-二磷酸钠盐

公司名称: Sigma-Aldrich
产品编号: A2754
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Platelet Migration and Bacterial Trapping Assay under Flow
Author:
Date:
2018-09-20
[Abstract]  Blood platelets are critical for hemostasis and thrombosis, but also play diverse roles during immune responses. We have recently reported that platelets migrate at sites of infection in vitro and in vivo. Importantly, platelets use their ability to migrate to collect and bundle fibrin (ogen)-bound bacteria accomplishing efficient intravascular bacterial trapping. Here, we describe a method that allows analyzing platelet migration in vitro, focusing on their ability to collect bacteria and trap bacteria under flow. [摘要]  血小板对于止血和血栓形成至关重要,但在免疫反应中也起着不同的作用。 我们最近报道了血小板在体外体外和体内感染部位迁移。 重要的是,血小板利用它们迁移的能力来收集和捆绑纤维蛋白(ogen)结合的细菌,从而实现有效的血管内细菌捕获。 在这里,我们描述了一种方法,允许分析血小板在体外的迁移,重点是它们收集细菌和捕获流动细菌的能力。

【背景】血小板是从巨核细胞释放的小的无核细胞片段,其存在于哺乳动物生物的骨髓内(Machlus和Italiano,2013)。大约7500亿血小板在人体血液中循环,不断扫描脉管系统以破坏内皮表面。在遇到内皮损伤时,血小板立即被招募在充分表征的事件级联中,包括初始血小板束缚和滚动,然后是血小板活化,粘附和扩散,最终导致纤维蛋白(ogen)依赖性聚集和随后的血栓收缩(Jackson, 2007)。血小板栓塞形成是生理性止血的主要步骤,但也是动脉粥样硬化斑块破裂后的病理性血栓形成,触发心肌梗塞或中风(Jackson,2011)。

除了在止血和血栓形成中的公认作用外,血小板还发展出多种免疫功能(Semple et al。,2011)。作为第一批招募炎症和感染部位的细胞,血小板在启动血管内免疫反应中起着重要作用(Wong et ...

Spectrophotometric Determination of Glutamine Synthetase Activity in Cultured Cells
Author:
Date:
2016-10-05
[Abstract]  Glutamine synthetase (GS), which catalyzes the conversion of glutamate and ammonia to glutamine, is widely distributed in animal tissues and cell culture lines. The importance of this enzyme is suggested by the fact that glutamine, the product of GS-catalyzed de novo synthesis reaction, is the most abundant free amino acid in blood (Smith and Wilmore, 1990). Glutamine is involved in many biological processes including serving as the nitrogen donor for biosynthesis, as an exchanger for the import of essential amino acids, as a means to detoxifying intracellular ammonia and glutamate, and as a ... [摘要]  谷氨酰胺合成酶(GS),其催化谷氨酸和氨转化成谷氨酰胺,广泛分布在动物组织和细胞培养系中。该酶的重要性通过谷氨酰胺,GS-催化的从头合成反应的产物,是血液中最丰富的游离氨基酸的事实提示(Smith和Wilmore,1990)。谷氨酰胺参与许多生物过程,包括作为生物合成的氮供体,作为输入必需氨基酸的交换剂,作为解毒细胞内氨和谷氨酸的手段,以及作为生物能量营养物来给三羧酸(TCA)周期(Bott等人,2015)。用于测定GS酶活性的方法依赖于其γ-谷氨酰转移酶反应,通过测量由谷氨酰胺和羟胺合成的γ-谷氨酰羟肟酸酯,以及反应产物与反应物的色谱分离(Deuel等人 。,1978)。 GS谷氨酰转移酶反应的概述可以在图1中找到。通过分光光度测定法在560nm的特定波长下使用酶标仪测量GS活性。该方法简单,并且具有与应用放射性标记的底物的那些方法相当的灵敏度。该修改的方法已经应用于在包括人乳腺上皮MCF10A细胞和鼠前B FL5.12细胞的培养细胞系中测定/测定GS活性,并且可以用于测量其他细胞系中的GS活性。 >


图1 。GS glutamyl ...

Determination of the H+-ATP Synthase and Hydrolytic Activities
Author:
Date:
2016-08-20
[Abstract]  The H+-ATP synthase of the inner mitochondrial membrane utilizes the proton gradient generated by the respiratory chain to synthesize ATP. Under depolarizing conditions, it can function in reverse by hydrolyzing ATP to generate a proton gradient. The protocols presented here allow the facile determination of both the synthetic and hydrolytic activities of the H+-ATP synthase in isolated mitochondria and in permeabilized mammalian cells. Since the protocol requires the isolation of polarized and well-coupled mitochondria, first we describe the protocol for mitochondrial ... [摘要]  内线粒体膜的H sup + -ATP合酶利用呼吸链产生的质子梯度来合成ATP。 在去极化条件下,它可以通过水解ATP产生质子梯度而反向作用。 本文提供的方案允许容易地测定分离的线粒体和透化的哺乳动物细胞中H sup + -ATP合酶的合成和水解活性。 由于该协议需要极化和良好耦合的线粒体的分离,首先我们描述线粒体从小鼠组织分离的协议。 第二,我们描述了用于测量ATP合成活性作为终点和在分离的线粒体和透化细胞中的动力学模式的方案。 最后,我们描述了用于测定酶在分离的线粒体中的ATP水解活性的方案。

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