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SorvallTM ST 40 Centrifuge Series

SorvallTM ST 40 Centrifuge Series

Company: Thermo Fisher Scientific
Catalog#: SorvallTM ST40R
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Pyocyanin Extraction and Quantitative Analysis in Swarming Pseudomonas aeruginosa
Author:
Date:
2016-12-05
[Abstract]  This protocol describes the quantification of pyocyanin extracted from swarming colonies of Pseudomonas aeruginosa. Pyocyanin is a secondary metabolite and a major virulence factor, whose production is inducible and varies highly under different growth conditions. The protocol is based on the earlier developed chloroform/HCl extraction of pyocyanin from liquid cultures (Frank and Demoss, 1959). Swarming colonies together with the agar they occupy are split into two halves. Pyocyanin is extracted from one of them. Cells are collected from the other half and used to quantify total protein yield and normalize the estimated corresponding pyocyanin quantities. [摘要]  该方案描述了从绿脓假单胞菌的群体群体提取的绿脓素的定量。绿脓素是次级代谢产物和主要毒力因子,其生产是可诱导的并且在不同生长条件下高度变化。该方案基于早先开发的氯仿/HCl从液体培养物中提取绿脓素(Frank和Demoss,1959)。聚集的菌落与它们所占据的琼脂一起分成两半。从它们之一提取绿脓菌素。从另一半收集细胞并用于定量总蛋白质产量并使估计的相应的绿脓素量标准化。
关键字:绿靛,Swarming,铜绿色,氯仿提取,毒力因子,运动性

[背景] 绿脓菌素是一种由人类病原体产生的蓝色氧化还原活性吩嗪色素。铜绿。绿脓菌素大量存在于感染有P的囊性纤维化患者的痰液中。 (Wilson等人,1988),并在病原体的毒力中起重要作用(Lau等人,2004)。绿脓菌素产生通过群体感应来调节,其涉及调节毒力基因的表达的信号分子的细胞密度依赖性合成(Fuqua等人,2001)。群集是响应于多种环境信号并使得细胞在半固体表面上的运动和宿主组织的建群而形成的复杂的共同行为。我们早先的观察表明,在液体培养物中的绿脓素生产。铜绿色与群体中的群体显着不同,可能是由于群体感应调节。因此,我们开发了一个协议,允许提取和定量的绿脓菌分泌的细胞成周围的琼脂分泌。所获得的量通过从群集细胞提取的总细胞蛋白质归一化。该方案不同于从固体琼脂表面(De ...

Human Blood Component Vaccinia Virus Neutralization Assay
Author:
Date:
2015-12-05
[Abstract]  Many therapeutic viruses, such as oncolytic viruses, vaccines, or gene therapy vectors, may be administered by the intravenous route to maximize their delivery to target tissues. Blood components, such as antibody, complement and blood cells (such as neutrophils, monocytes, T cells, B cells or platelets) may result in viral neutralization and therefore reduce the therapeutic efficacy. This protocol will describe an in vitro assay by which to test the interaction of viruses with blood components. The effect of various factors can be isolated through fractionation. While whole blood can offer the most physiologically relevant snapshot, plasma can investigate the effects of antibody in concert with complement, and heat inactivated plasma will interrogate the effect of antibody ... [摘要]  许多治疗性病毒,例如溶瘤病毒,疫苗或基因治疗载体,可以通过静脉内途径施用以最大化它们向靶组织的递送。 血液组分,例如抗体,补体和血细胞(例如嗜中性粒细胞,单核细胞,T细胞,B细胞或血小板)可导致病毒中和并因此降低治疗功效。 该方案将描述用于测试病毒与血液组分的相互作用的体外测定。 各种因素的影响可以通过分馏分离。 尽管全血可以提供最具生理学相关性的快照,但血浆可以研究抗体与补体一起的作用,而热灭活的血浆将会询问单独的抗体的作用。

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