用于评估货物向溶酶体转运的培养细胞中DQ-红色牛血清白蛋白转运分析
DQ-Red BSA Trafficking Assay in Cultured Cells to Assess Cargo Delivery to Lysosomes.
作者信息
Marwaha Rituraj, Sharma Mahak
机构信息
Department of Biological Sciences, Indian Institute of Science Education and Research Mohali (IISERM), Punjab, India.
出版信息
Bio Protoc. 2017 Oct 5;7(19). doi: 10.21769/BioProtoc.2571.
Abstract
Lysosomes are the terminal end of the endocytic pathway having acidic environment required for active hydrolases that degrade the cargo delivered to these compartments. This process of cargo delivery and degradation by endo-lysosomes is a tightly regulated process and important for maintaining cellular homeostasis. Cargos like EGF (Epidermal Growth Factor), Dil-LDL (3,3'-Dioctadecylindocarbocyanine-Low Density Lipoprotein), Dextran, DQ-BSA (Dye Quenched-Bovine Serum Albumin) are routinely used by researchers to analyze the role of various proteins in endocytic pathway. Trafficking of DQ-BSA in cells depleted of or over-expressing the gene of interest is a useful assay for identifying the role of various proteins in endocytic trafficking pathway. The protocol describes the DQ-Red BSA trafficking assay that can be used to study endocytic trafficking in various cell types.
摘要
溶酶体是内吞途径的终点，具有活跃水解酶所需的酸性环境，这些水解酶可降解输送至这些区室的货物。内吞溶酶体进行货物递送和降解的这一过程受到严格调控，对维持细胞内稳态至关重要。诸如表皮生长因子（EGF）、3,3'-二辛基吲哚碳菁标记的低密度脂蛋白（Dil-LDL）、葡聚糖、染料淬灭牛血清白蛋白（DQ-BSA）等货物常被研究人员用于分析各种蛋白质在内吞途径中的作用。在缺乏或过表达目标基因的细胞中检测DQ-BSA的运输情况，是一种用于确定各种蛋白质在内吞运输途径中作用的有效检测方法。本实验方案描述了可用于研究多种细胞类型内吞运输的DQ-红色牛血清白蛋白运输检测方法。
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