Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody.
Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody.
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DOI：
10.1016/j.xpro.2023.102271
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发表时间：
2023-06-07
期刊：
影响因子：
--
通讯作者：
Zhao, Na
中科院分区：
文献类型：
--
作者：
O'Leary, Justin;Raulin, Ana-Caroline;Li, Zonghua;Martens, Yuka;Inoue, Yasuteru;Strickland, Michael R.;Han, Xianlin;Holtzman, David M.;Bu, Guojun;Zhao, Na
载脂蛋白E蛋白（apoE）赋予阿尔茨海默病的不同风险取决于表达的同种型。在这里，我们提出了一个为期2天的免疫沉淀协议，使用HJ15.4单克隆apoE抗体的下拉天然apoE颗粒。我们描述了通过永生化星形胶质细胞培养和HJ15.4抗体珠偶联apoE颗粒下拉，洗脱和表征apoE生产的主要步骤。该方案可用于从多个模型系统或人类生物标本中分离天然apoE颗粒。APOE 3/3永生化星形胶质细胞培养条件和条件培养基收集分离天然apoE脂蛋白颗粒的详细两天免疫沉淀方案天然apoE脂蛋白颗粒大小分布和脂质谱的表征方案适用于多种apoE脂蛋白颗粒来源和下游测定出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和伦理指南。载脂蛋白E蛋白（apoE）赋予阿尔茨海默病的不同风险取决于表达的同种型。在这里，我们提出了一个为期2天的免疫沉淀协议，使用HJ15.4单克隆apoE抗体的下拉天然apoE颗粒。我们描述了通过永生化星形胶质细胞培养和HJ15.4抗体珠偶联apoE颗粒下拉，洗脱和表征apoE生产的主要步骤。该方案可用于从多个模型系统或人类生物标本中分离天然apoE颗粒。
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. APOE3/3 immortalized astrocyte culture conditions and conditioned media collection Detailed two-day immunoprecipitation protocol isolating native apoE lipoprotein particles Characterization of native apoE lipoprotein particle size distribution and lipid profile Protocol applicable to multiple apoE lipoprotein particle sources and downstream assays Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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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影响因子：
4.2
作者：
DeMattos, RB;Brendza, RP;Holtzman, DM
通讯作者：
Holtzman, DM
影响因子：
12.7
作者：
Zhao J;Lu W;Ren Y;Fu Y;Martens YA;Shue F;Davis MD;Wang X;Chen K;Li F;Liu CC;Graff-Radford NR;Wszolek ZK;Younkin SG;Brafman DA;Ertekin-Taner N;Asmann YW;Dickson DW;Xu Z;Pan M;Han X;Kanekiyo T;Bu G
通讯作者：
Bu G
影响因子：
15.9
作者：
Liao, Fan;Li, Aimin;Holtzman, David M.
通讯作者：
Holtzman, David M.
影响因子：
10.6
作者：
Zhao N;Liu CC;Qiao W;Bu G
通讯作者：
Bu G
影响因子：
16.2
作者：
Martens, Yuka A.;Zhao, Na;Liu, Chia-Chen;Kanekiyo, Takahisa;Yang, Austin J.;Goate, Alison M.;Holtzman, David M.;Bu, Guojun
通讯作者：
Bu, Guojun