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10 ml sterile, disposable pipettes

Company: Thermo Fisher Scientific
Catalog#: 4488
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A Quantitative Single-cell Flow Cytometry Assay for Retrograde Membrane Trafficking Using Engineered Cholera Toxin
Author:
Date:
2020-08-05
[Abstract]  The organization and distribution of proteins, lipids, and nucleic acids in eukaryotic cells is an essential process for cell function. Retrograde trafficking from the plasma membrane to the Golgi and endoplasmic reticulum can greatly modify cell membrane composition and intracellular protein dynamics, and thus typifies a key sorting step. However, methods to efficiently quantify the extent or kinetics of these events are currently limited. Here, we describe a novel quantitative and effectively real-time single-cell flow cytometry assay to directly measure retrograde membrane transport. The assay takes advantage of the well-known retrograde trafficking of cholera toxin engineered with split-fluorescent proteins to generate novel tools for immediate monitoring of intracellular trafficking. ... [摘要]  [摘要]蛋白质、脂类和核酸在真核细胞中的组织和分布是细胞功能的重要过程。从质膜到高尔基体和内质网的逆向运输可以极大地改变细胞膜的组成和细胞内蛋白质的动态变化,因此是一个关键的分选步骤。然而,有效量化这些事件的程度或动力学的方法目前是有限的。在这里,我们描述了一种新的定量和有效的实时单细胞流式细胞术检测直接测量逆行膜转运。这项检测利用了众所周知的霍乱毒素逆行转移的特性,利用裂解荧光蛋白产生了新的工具,用于即时监测细胞内的转移。这种方法将大大扩展研究细胞内膜转运的生物学基础,以及细胞膜转运系统如何适应不同细胞类型和细胞状态的生理需要。

[背景]所有的真核细胞都依赖于它们动态地将分子分类和分离到膜结合的亚细胞器中的能力,以便组织和分配到细胞的特定区域。在这一过程中的一个重要步骤是早期分类内体和跨高尔基网络(TGN)。在从分选内体产生的其他贩运途径中,通过分泌途径到TGN的逆行贩运是一个关键的分选步骤(Johannes和Popoff,2008)。细胞膜蛋白和脂类发生逆向转运。从TGN到内质网(ER)的进一步逆行贩运可通过一些质膜脂质完成,并可巧妙地由几种细菌毒素和病毒协同作用而致病(Cho等人,2012年;Personnic等人,2016年;Williams和Tsai,2016年)。

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Retroviral Capsid Core Stability Assay
Author:
Date:
2018-09-20
[Abstract]  Structural stability of the capsid core is a critical parameter for the productive infection of a cell by a retrovirus. Compromised stability can lead to premature core disassembly, exposure of replication intermediates to cytosolic nucleic acid sensors that can trigger innate antiviral responses, and failure to integrate the proviral genome into the host DNA. Thus, core stability is a critical feature of viral replicative fitness. While there are several well-described techniques to assess viral capsid core stability, most are generally time and labor intensive. Recently, our group compared the relative stability of murine leukemia virus capsid cores using an in vitro detergent-based approach combined with ultracentrifugation against the popular fate of capsid assay. We found ... [摘要]  衣壳核心的结构稳定性是逆转录病毒对细胞的生产性感染的关键参数。受损的稳定性可导致核心过早解体,将复制中间体暴露于细胞溶质核酸传感器,其可触发先天的抗病毒反应,并且不能将原病毒基因组整合到宿主DNA中。因此,核心稳定性是病毒复制适应性的关键特征。虽然有几种充分描述的技术来评估病毒衣壳核心稳定性,但大多数通常是时间和劳动密集型的。最近,我们小组使用基于体外洗涤剂的方法结合超速离心法对衣壳测定的流行命运比较了鼠白血病病毒衣壳核心的相对稳定性。我们发现两种方法都得出了类似的结论,尽管第一种方法是比较显示核心稳定性差异的病毒突变体时评估相对衣壳核心稳定性的一种非常简单和快速的方法。

【背景】逆转录病毒已经进化出复制周期,在规避宿主抗病毒反应方面表现优异。逆转录病毒开发的一种策略是将其复制中间体与胞质核酸传感器如cGAS,TREX1,IFI203和DDX41屏蔽起来(Yan et al。,2010; Gao et al。,2013; Lahaye et al。,2013; Stavrou et al。,2015)。在复制过程中,逆转录病毒在细胞质中产生RNA-DNA杂合体和未甲基化的双链前病毒DNA,这是先天免疫传感器的常见靶标(Yan et al。,2010; Gao et al。,2013; Lahaye et ...

Qualitative in vivo Bioluminescence Imaging
Author:
Date:
2018-09-20
[Abstract]  Bioluminescence imaging (BLI) technology is an advanced method of carrying out molecular imaging on live laboratory animals in vivo. This powerful technique is widely-used in studying a variety of biological processes, and it has been an ideal tool in exploring tumor growth and metastatic spread in real-time. This technique ensures the optimal use of laboratory animal resources, particularly the ethical principle of reduction in animal use, given its non-invasive nature, ensuring that ongoing biological processes can be studied over time in the same animal, without the need to euthanize groups of mice at specific time points. In this protocol, the luciferase imaging technique was developed to study the effect of co-inoculating pericytes (contractile, αSMA+ mesenchymal ... [摘要]  生物发光成像(BLI)技术是一种在体内实验室动物上进行分子成像的先进方法。 这种强大的技术广泛应用于研究各种生物过程,是实时探索肿瘤生长和转移扩散的理想工具。 该技术确保实验室动物资源的最佳利用,特别是减少动物使用的伦理原则,考虑到其非侵入性,确保可以在同一动物中随时间研究正在进行的生物过程,而无需安乐死 小鼠在特定的时间点。 在该方案中,开发了荧光素酶成像技术以研究共同接种周细胞(收缩性,αSMA + 间充质干细胞样细胞,位于微血管内的细胞)对生长和转移性扩散的影响。 卵巢癌使用侵袭性卵巢癌细胞系-OVCAR-5-作为例子。

【背景】生物发光成像(BLI)的原理是基于相对简单的生化过程的发光特性,即,荧光素酶介导的分子底物荧光素氧化产生光。在癌症研究中,BLI是一种流行的工具(Contag et ...

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