突变型亨廷顿蛋白在Neuro2A细胞和大鼠原代皮质神经元中的分泌
Mutant Huntingtin Secretion in Neuro2A Cells and Rat Primary Cortical Neurons.
作者信息
Trajkovic Katarina, Jeong Hyunkyung, Krainc Dimitri
机构信息
Department of Neurology, Northwestern University Feinberg School of Medicine, Chicago, Illinois, USA.
出版信息
Bio Protoc. 2018 Jan 5;8(1). doi: 10.21769/BioProtoc.2675.
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.
由于细胞分泌的蛋白质丰度低，且细胞培养基中存在大量干扰性的牛血清白蛋白（BSA），对这些蛋白质进行定量分析具有挑战性。我们通过蛋白质印迹法建立了检测从稳定表达突变亨廷顿蛋白（mHtt）的Neuro2A细胞系和大鼠原代皮质神经元分泌的mHtt的检测方法。我们的方案基于在保持细胞活力和分泌潜能的同时减少培养基中BSA的量，并在分析前通过超滤浓缩培养基。
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