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

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

doi:10.1016/j.xpro.2023.102271

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

STAR Protoc. 2023 Jun 7;4(2):102271. doi: 10.1016/j.xpro.2023.102271. Online ahead of print.

Authors

Affiliations

¹ Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA. Electronic address: oleary.justin@mayo.edu.
² Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA.
³ Department of Neurology, Hope Center for Neurological Disorders, Knight Alzheimer's Disease Research Center, Washington University in St. Louis, St. Louis, MO, 63110, USA.
⁴ Department of Medicine, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA; Barshop Institute for Longevity and Aging Studies, University of Texas Health Science Center at San Antonio, San Antonio, TX 78229, USA.
⁵ Department of Neuroscience, Mayo Clinic, Jacksonville, FL 32224, USA. Electronic address: zhao.na@mayo.edu.

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.

Keywords: Cell Biology; Molecular Biology; Neuroscience.

Abstract

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