Details

Autor(en) / Beteiligte

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

Titel

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

Ist Teil von

• STAR protocols, 2023-06, Vol.4 (2), p.102271-102271, Article 102271

Ort / Verlag

United States: Elsevier Inc

Erscheinungsjahr

2023

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• 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. [Display omitted] •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.