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Calcium chloride dihydrate

氯化钙二水合物

Company: Sigma-Aldrich
Catalog#: C3306
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Combining Gel Retardation and Footprinting to Determine Protein-DNA Interactions of Specific and/or Less Stable Complexes
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Date:
2020-12-05
[Abstract]  

DNA footprinting is a classic technique to investigate protein-DNA interactions. However, traditional footprinting protocols can be unsuccessful or difficult to interpret if the binding of the protein to the DNA is weak, the protein has a fast off-rate, or if several different protein-DNA complexes are formed. Our protocol differs from traditional footprinting protocols, because it provides a method to isolate the protein-DNA complex from a native gel after treatment with the footprinting agent, thus removing the bound DNA from the free DNA or other protein-DNA complexes. The DNA is then extracted from the isolated complex before electrophoresis on a sequencing gel to determine the footprinting pattern. This analysis provides a possible solution for those who have been unable to use

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[摘要]  [摘要] DNA足迹是研究蛋白质-DNA相互作用的经典技术。但是,如果蛋白质与DNA的结合较弱,蛋白质的脱落速率较快,或者形成了几种不同的蛋白质-DNA复合物,则传统的足迹方案可能会失败或难以解释。我们的协议不同于传统的足迹协议,因为它提供了一种在使用足迹剂处理后从天然凝胶中分离蛋白质-DNA复合物的方法,从而从游离DNA或其他蛋白质-DNA复合物中去除了结合的DNA。然后从分离的复合物中提取DNA,然后在测序凝胶上电泳以确定印迹模式。该分析为无法使用传统足迹法确定蛋白质与DNA接触的人提供了可能的解决方案。

[背景]核酸酶/化学足迹是一个典型的方法来探测蛋白质-DNA相互作用(腊士和施米茨,1978;萨瑟-德怀特和Gralla,1989;汉普等人,2007) ...

Flow Cytometry Analysis and Fluorescence-activated Cell Sorting of Myeloid Cells from Lung and Bronchoalveolar Lavage Samples from Mycobacterium tuberculosis-infected Mice
Author:
Date:
2020-05-20
[Abstract]  Mycobacterium tuberculosis (Mtb) is transmitted by aerosol and can cause serious bacterial infection in the lung that can be fatal if left untreated. Mtb is now the leading cause of death worldwide by an infectious agent. Characterizing the early events of in vivo infection following aerosol challenge is critical for understanding how innate immune cells respond to infection but is technically challenging due to the small number of bacteria that initially infect the lung. Previous studies either evaluated Mtb-infected cells at later stages of infection when the number of bacteria in the lung is much higher or used in vitro model systems to assess the response of myeloid cells to Mtb. Here, we describe a method that uses fluorescent bacteria, a high-dose aerosol ... [摘要]  [摘要 ] 结核分枝杆菌(Mtb)通过气溶胶传播,可引起严重的肺部细菌感染,如果不及时治疗,可能致命。Mtb现在已成为全球传染病致死的主要原因。表征气溶胶激发后体内感染的早期事件对于了解先天免疫细胞如何对感染做出反应至关重要,但由于最初会感染肺的细菌数量少,因此在技术上具有挑战性。先前的研究或者在肺部细菌数量高得多时在感染后期评估Mtb感染的细胞,或者在体外使用 评估骨髓细胞对Mtb反应的模型系统。在这里,我们介绍一种使用荧光细菌,大剂量气溶胶感染模型和流式细胞术跟踪气溶胶感染和荧光激活细胞分选(FACS)之后立即分离肺中Mtb感染细胞的方法,以分离幼稚的旁观者,和Mtb感染的细胞用于下游应用,包括RNA测序。该协议提供了在肺环境中监视Mtb感染和细胞特异性反应的能力,已知该环境可调节常驻和募集人群的功能。使用此协议,我们发现肺泡巨噬细胞通过上调受转录因子Nrf2调节并有害于细菌早期控制的细胞保护性转录反应,在体内对Mtb感染作出反应。

[背景 ] 气溶胶传播是结核分枝杆菌(Mtb)感染自然周期的关键组成部分,有助于细菌的毒性并导致其在肺部的独特感染模式(North ,1995;Riley 等,1995)。 ; Pai et ...

Pentylenetetrazole (PTZ)-induced Convulsion Assay to Determine GABAergic Defects in Caenorhabditis elegans
Author:
Date:
2018-09-05
[Abstract]  Pentylenetetrazole (PTZ) is a GABAA receptor antagonist and is used to monitor presynaptic defects in the release of the inhibitory neurotransmitter GABA. PTZ is a competitive inhibitor of GABA, and prevents binding of GABA on the GABAA receptors present on the surface of muscle. In the absence of GABA binding, the excitatory to inhibitory signal ratio increases resulting in a convulsive phenotype. This assay provides a fast and reliable method to detect presynaptic defects in GABAergic synaptic transmission. The assay is based on correlating the extent of convulsions with the degree of presynaptic GABA release defects. [摘要]  戊四唑(PTZ)是GABA A 受体拮抗剂,用于监测抑制性神经递质GABA释放中的突触前缺陷。 PTZ是GABA的竞争性抑制剂,并且阻止GABA与肌肉表面上存在的GABA A 受体结合。 在没有GABA结合的情况下,抑制信号比的兴奋性增加导致惊厥表型。 该试验提供了一种快速可靠的方法来检测GABA能突触传递中的突触前缺陷。 该测定基于将惊厥的程度与突触前GABA释放缺陷的程度相关联。

【背景】突触是介导突触传递的不对称细胞间连接。它们构成了能够执行复杂行为的大脑电路的基本单元。动作电位的到达通过电压门控钙通道引起钙流入,导致质膜的去极化。在这种突触前去极化后,突触囊泡与质膜融合,通过高度控制的过程将其内容物释放到突触间隙中(Sudhof,1995; Sudhof,2004)。

运动是 Caenorhabditis elegans 中突出的行为输出。产生协调的背腹窦弯曲的神经回路允许 C中的正常运动。线虫。运动行为在多个层面上进行协调,并且涉及由中间神经元处理的多个感觉神经元整合多种感觉线索,并最终在神经肌肉接头(NMJ)处直接变化(de Bono和Maricq,2005; Bargmann,2012)。 C.线虫 NMJ有三个主要组成部分;兴奋性胆碱能运动神经元,抑制性GABA能神经运动神经元和突触后肌细胞(White et ...

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