| Mutant Huntingtin Secretion in Neuro2A Cells and Rat Primary Cortical Neurons
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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 ...
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| In vitro Drug Susceptibility Assay for HBV Using Southern Blotting
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Author:
Date:
2015-04-20
[Abstract] Antiviral agents for the suppression of hepatitis B virus (HBV) have been used for treating chronic hepatitis B. However, the emergence of drug-resistant HBV is still a major problem for antiviral treatment. To identify and characterize the drug-resistant HBV, the construction of HBV replicon and in vitro drug susceptibility assay are essential. Here we describe the experimental methods to study drug-resistant HBV.
[摘要] 用于抑制乙型肝炎病毒（HBV）的抗病毒剂已经用于治疗慢性乙型肝炎。然而，耐药性HBV的出现仍然是抗病毒治疗的主要问题。 为了鉴定和表征耐药性HBV，构建HBV复制子和体外药物敏感性测定是必需的。 在这里我们描述研究耐药性HBV的实验方法。
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