Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody

利用 HJ15.4 单克隆抗体高效分离天然载脂蛋白 E 颗粒的标准化免疫沉淀方案

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作者:Justin O'Leary, Ana-Caroline Raulin, Zonghua Li, Yuka Martens, Yasuteru Inoue, Michael R Strickland, Xianlin Han, David M Holtzman, Guojun Bu, Na Zhao

Abstract

The apolipoprotein E protein (apoE) confers differential risk for Alzheimer's disease depending on which isoforms are expressed. Here, we present a 2-day immunoprecipitation protocol using the HJ15.4 monoclonal apoE antibody for the pull-down of native apoE particles. We describe major steps for apoE production via immortalized astrocyte culture and HJ15.4 antibody bead coupling for apoE particle pull-down, elution, and characterization. This protocol could be used to isolate native apoE particles from multiple model systems or human biospecimens.

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