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Silicone grease - SILICAID 1010

Company: aidchim Ltd
Catalog#: H3375
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HIVGKO: A Tool to Assess HIV-1 Latency Reversal Agents in Human Primary CD4+ T Cells
Author:
Date:
2018-10-20
[Abstract]  While able to suppress HIV replication in HIV infected individuals, combination antiretroviral therapy (ART) fails to eliminate viral latent reservoir, which consists in integrated transcriptional silenced HIV provirus. So far, identification of latently-infected cells has relied on activating cells to induce expression of HIV proteins which can then be detected. Unfortunately, this activation significantly changed the cell phenotype. We developed a novel HIV reporter, named HIVGKO, that allows the purification of latently-infected cells in absence of reactivation. Indeed, latent cells can be identified by expression of the EF1a-driven mKO2 and lack of expression of the LTR-driven csGFP. This protocol can be used to study the effectiveness of LRAs (Latency Reversal Agents) in ... [摘要]  虽然能够抑制HIV感染个体中的HIV复制,但联合抗逆转录病毒疗法(ART)无法消除病毒潜伏性储库,其包含整合的转录沉默的HIV原病毒。 到目前为止,潜伏感染细胞的鉴定依赖于激活细胞以诱导HIV蛋白的表达,然后可以检测到这些蛋白的表达。 不幸的是,这种激活显着改变了细胞表型。 我们开发了一种名为HIV GKO 的新型HIV报告基因,可以在没有再激活的情况下纯化潜伏感染的细胞。 实际上,可以通过EF1a驱动的mKO2的表达和LTR驱动的csGFP的缺乏表达来鉴定潜伏细胞。 该方案可用于研究LCA(潜伏期逆转剂)在原代细胞中重新激活潜伏HIV的有效性。

【背景】新版双标记病毒(HIV GKO )含有5'LTR中HIV-1启动子控制下的密码子转换eGFP(csGFP)和一种独特的无关荧光蛋白 mKO2在细胞延伸因子αα启动子(EF1α)的控制下。 当使用与遗传相关的荧光蛋白时,由于重组问题,在这些报道分子中使用不相关的荧光蛋白是很重要的。 因此,生产性感染的细胞主要是csGFP + mKO2 + (有些可能只是GFP + ),而潜伏感染的细胞是csGFP - mKO2 + 。 流式细胞仪如分拣机AriaII允许纯化纯感染人群(生产性,潜伏性和/或未感染),而分析仪LSRII允许评估HIV GKO LTR的转录激活。 感染后的时间很短。

Isolation of Chromatin-bound Proteins from Subcellular Fractions for Biochemical Analysis
Author:
Date:
2018-10-05
[Abstract]  Shuttling of proteins between different cellular compartments controls their proteostasis and can contribute in some cases to regulate their activity. Biochemical analysis of chromatin-bound proteins, such as transcription factors, is often difficult because of their low yield and due to the interference from nucleic acids. This protocol describes a method to efficiently fractionate cells combined with a mechanical (i.e., sonication) or an enzymatic treatment (i.e., benzonase) that facilitates analysis of chromatin-bound protein extracts by Western blot analysis or by protein pull-down assays. This approach can be valuable to enrich a particular protein within a particular subcellular fraction either to study specific post-translational modification patterns or to ... [摘要]  在不同细胞区室之间穿梭蛋白质控制它们的蛋白质稳态,并且在某些情况下可以有助于调节它们的活性。 染色质结合蛋白(例如转录因子)的生化分析通常是困难的,因为它们的产率低并且由于核酸的干扰。 该协议描述了一种有效分离细胞的方法,结合机械(即,超声处理)或酶处理(即,benzonase),有助于分析染色质结合蛋白提取物 通过蛋白质印迹分析或蛋白质下拉分析。 该方法对于富集特定亚细胞级分内的特定蛋白质以研究特定的翻译后修饰模式或鉴定特定的蛋白质 - 蛋白质相互作用可能是有价值的。
【背景】许多染色质结合蛋白的活性和翻译后调节研究很少,因为在分离它们进行生化分析时存在技术困难。这甚至是转录因子的情况,例如基本的螺旋 - 环 - 螺旋(bHLH)转录因子,其通常在组织或细胞模型中具有稀缺的时间和空间表达模式(Dennis 等。,2018)。当生物材料的量成为研究分子途径的障碍时,协议细化有助于解除技术限制(Gillotin和Guillemot,2016)。在我们最近的研究中,我们努力了解神经元bHLH转录因子Ascl1的蛋白水解是如何在神经元分化的细胞模型中调节的(Gillotin et ...

Detection of Cell Death in Planarians
Author:
Date:
2018-10-05
[Abstract]  Planarians are freshwater flatworms, well known for their ability to regenerate a complete organism from any piece of their body. Furthermore, planarians are constantly growing and degrowing throughout their lives, maintaining a functional and proportioned body. These properties rely on the presence of a population of adult stem cells and on the tight control of their cell renewal, which is based on the balance between the proliferation of new cells and their differentiation, and the death of unnecessary cells. Due to the importance of these two processes in planarian biology, over the years, researchers have optimized molecular techniques to detect both cell proliferation and cell death in planarians. Here, we present the two main protocols currently used for cell death detection and ... [摘要]  涡虫是淡水扁虫,因其能够从身体的任何部分再生完整的有机体而闻名。 此外,涡虫在其一生中不断生长和去除,保持功能和比例的身体。 这些特性依赖于成体干细胞群的存在以及对细胞更新的严格控制,其基于新细胞增殖与其分化之间的平衡以及不必要细胞的死亡。 由于这两个过程在涡虫生物学中的重要性,多年来,研究人员已经优化了分子技术,以检测涡虫中的细胞增殖和细胞死亡。 在这里,我们提出了目前用于细胞死亡检测和量化的两种主要方案:Caspase-3活性定量和TUNEL分析。
【背景】成体生物体中的细胞更新是基于三个过程的复杂机制:(a)通过细胞死亡消除选定的细胞; (b)通过细胞分裂取代已消除的细胞,通常涉及成体干细胞及其后代; (c)新生细胞的分化及其与先前存在的组织的整合(Pellettieri和Sanchez ...

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