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Boric acid

硼酸

公司名称: Sigma-Aldrich
产品编号: B7660
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Mutant Huntingtin Secretion in Neuro2A Cells and Rat Primary Cortical Neurons
Author:
Date:
2018-01-05
[Abstract]  Quantitative analysis of proteins secreted from the cells poses a challenge due to their low abundance and the interfering presence of a large amount of bovine serum albumin (BSA) in the cell culture media. We established assays for detection of mutant huntingtin (mHtt) secreted from Neuro2A cell line stably expressing mHtt and rat primary cortical neurons by Western blotting. Our protocol is based on reducing the amounts of BSA in the media while maintaining cell viability and secretory potential, and concentrating the media prior to analysis by means of ultrafiltration. [摘要]  由细胞分泌的蛋白质的定量分析由于它们的丰度低和在细胞培养基中干扰大量牛血清白蛋白(BSA)的存在而提出挑战。 我们建立检测突变亨廷顿蛋白(mHtt)检测分泌Neuro2A细胞系稳定表达mHtt和大鼠原代皮层神经元蛋白质印迹。 我们的方案是基于降低培养基中BSA的量,同时保持细胞活力和分泌潜能,并在通过超滤分析之前浓缩培养基。


【背景】许多蛋白质通过各种分泌途径从细胞分泌到细胞外环境中。这些途径包括在ER-高尔基体 - 质膜途径之后的常规分泌途径(Lee等,2004)和多个非常规途径,例如溶酶体胞吐作用,穿过质膜的易位和外泌体以及胞外体释放(Zhang和Schekman,2013)。为了研究这些途径,经常需要分析培养细胞分泌的蛋白质进入培养基。蛋白质可以游离形式分泌,也可以与胞膜结构如胞外体和外泌体结合(Zhang and ...

Neuron Culture from Mouse Superior Cervical Ganglion
Author:
Date:
2014-01-20
[Abstract]  The rodent superior cervical ganglion (SCG) is a useful and readily accessible source of neurons for studying the mechanisms of sympathetic nervous system (SNS) development and growth in vitro. The sympathetic nervous system (SNS) of early postnatal animals undergoes a great deal of remodeling and development; thus, neurons taken from mice at this age are primed to re-grow and establish synaptic connections after in situ removal. The stereotypic location and size of the SCG make it ideal for rapid isolation and dissociation. The protocol described here details the ... [摘要]  啮齿动物上级子宫颈神经节(SCG)是一种有用且易于获取的神经元来源,用于研究交感神经系统(SNS)体外发育和生长的机制。 早期出生后动物的交感神经系统(SNS)经历了大量的重塑和发育; 因此,在这个年龄的小鼠中取出的神经元被引发再生长并在原位移除后建立突触连接。 SCG的刻板位置和尺寸使其成为快速隔离和解离的理想选择。 这里描述的方案详述了SCG神经元的解剖,培养和分化的要求。 该方案适用于从晚期胚胎妊娠至大约出生后第3天培养神经元。下面讨论的培养技术利用玻璃盖玻片进行固定细胞的显微镜检查。

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