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Potassium chloride

氯化钾

Company: Sigma-Aldrich
Catalog#: P9541
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Sacral Spinal Cord Transection and Isolated Sacral Cord Preparation to Study Chronic Spinal Cord Injury in Adult Mice
Author:
Date:
2018-04-05
[Abstract]  Spinal cord injury (SCI) is characterized by multiple sensory/motor impairments that arise from different underlying neural mechanisms. Linking specific sensory/motor impairments to neural mechanism is limited by a lack of direct experimental access to these neural circuits. Here, we describe an experimental model which addresses this shortcoming. We generated a mouse model of chronic spinal cord injury that reliably reproduces spasticity observed after SCI, while at the same time allows study of motor impairments in vivo and in an in vitro preparation of the spinal cord. The model allows for the combination of mouse genetics in in vitro and in vivo conditions with advanced imaging, behavioral analysis, and detailed electrophysiology, techniques which ... [摘要]  脊髓损伤(SCI)的特点是由不同的潜在神经机制引起的多种感觉/运动损伤。 将特定的感觉/运动障碍与神经机制联系起来受限于缺乏对这些神经回路的直接实验访问。 在这里,我们描述了一个解决这个缺点的实验模型。 我们产生了一种慢性脊髓损伤的小鼠模型,其可靠地再现SCI后观察到的痉挛状态,同时允许研究运动损伤体内和体外制剂 的脊髓。 该模型允许将体外和体内条件下的小鼠遗传与高级成像,行为分析和详细的电生理学技术相结合,这些技术在常规SCI中不易应用 楷模。


【背景】脊髓损伤导致了毁灭性的感官 - ...

Tracking Lipid Transfer by Fatty Acid Isotopolog Profiling from Host Plants to Arbuscular Mycorrhiza Fungi
Author:
Date:
2018-04-05
[Abstract]  Lipid transfer from host plants to arbuscular mycorrhiza fungi was hypothesized for several years because sequenced arbuscular mycorrhiza fungal genomes lack genes encoding cytosolic fatty acid synthase (Wewer et al., 2014; Rich et al., 2017). It was finally shown by two independent experimental approaches (Jiang et al., 2017; Keymer et al., 2017; Luginbuehl et al., 2017). One approach used a technique called isotopolog profiling (Keymer et al., 2017). Isotopologs are molecules, which differ only in their isotopic composition. For isotopolog profiling an organism is fed with a heavy isotope labelled precursor metabolite. Subsequently, the labelled isotopolog composition of metabolic products is analysed via mass spectrometry. The ... [摘要]  因为测序的丛枝菌根真菌基因组缺乏编码胞质脂肪酸合酶的基因(Wewer等人,2014; Rich等人,2014),因此假定脂质从宿主植物转移到丛枝菌根真菌数年。 / em>,2017)。最终通过两种独立的实验方法(Jiang等人,2017; Keymer等人,2017; Luginbuehl等人, ), 2017年)。一种方法使用称为同位素体谱分析的技术(Keymer等人,2017)。同位素体是分子,它们的同位素组成不同。对于同位素生物学分析,生物体被喂以重同位素标记的前体代谢物。随后,通过质谱分析代谢产物的标记同位素组成。检测到的目标代谢物的同位素体模式产生关于代谢途径和通量的信息(Ahmed et al。,2014)。以下协议描述了一个实验装置,该装置能够在由丛枝菌根真菌及其相关真菌胞外菌丝体定植的植物根中分离出脂肪酸的单独同位素分布图,以阐明两种共生生物之间的通量。我们预测,如果两种相互作用的生物体可以物理分离,则该策略还可以用于研究其他生物体之间的代谢物通量。

【背景】丛枝菌根真菌是生物营养生物。因此,它们不能独立栽培,而是依靠与寄主植物的相互作用来维持生命并完成它们的生命周期。这一特征使得研究两种共生生物,特别是单独的真菌具有挑战性。

为了培养,处理和收获与宿主根分开的真菌,开发了2室培养皿系统,并将其用于以前工作中的标记研究(Bécard和Fortin,1988; ...

Purification of RNA Mango Tagged Native RNA-protein Complexes from Cellular Extracts Using TO1-Desthiobiotin Fluorophore Ligand
Author:
Date:
2018-04-05
[Abstract]  A native purification strategy using RNA Mango for RNA based purification of RNA-protein complexes is described. The RNA Mango aptamer is first genetically engineered into the RNA of interest. RNA Mango containing complexes obtained from cleared cellular native extracts are then immobilized onto TO1-Desthiobiotin saturated streptavidin agarose beads. The beads are washed to remove non-specific complexes and then the RNA Mango containing complexes are eluted by the addition of free biotin to the beads. Since the eluted complexes are native and fluorescent, a second purification step such as size exclusion chromatography can easily be added and the purified complexes tracked by monitoring fluorescence. The high purity native complexes resulting from this two-step purification strategy can ... [摘要]  描述了使用RNA Mango进行RNA-蛋白质复合物的RNA纯化的天然纯化策略。 RNA芒果适体首先被基因工程改造成感兴趣的RNA。 然后将从清除的细胞天然提取物获得的含有RNA复合物的复合物固定在TO1-Desthiobiotin饱和的链霉亲和素琼脂糖珠上。 洗涤珠粒以去除非特异性复合物,然后通过向珠粒中加入游离生物素来洗脱含RNA芒果的复合物。 由于洗脱的复合物是天然的和荧光的,所以可以容易地添加第二纯化步骤如尺寸排阻色谱,并且通过监测荧光追踪纯化的复合物。 通过这种两步纯化策略产生的高纯度天然复合物可以用于进一步的生物化学表征。

【背景】目前的RNA标签受限于诸如差K ,大尺寸,潜在的生物学干扰或缺乏固有荧光的限制(Panchapakesan等人, 2015年)。 RNA芒果很小,可以简单地整合到茎环结构中,特别是GNRA tetraloops中,它具有生物耐受性,并且最重要的是对其噻唑橙基(TO1)配体TO1-Desthiobiotin(TO1-Dtb)具有高亲和力。 ...

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