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

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

Neuroscience

Research output: Contribution to journal › Article › peer-review

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.

Original language	English (US)
Article number	102271
Journal	STAR Protocols
Volume	4
Issue number	2
DOIs	https://doi.org/10.1016/j.xpro.2023.102271
State	Published - Jun 16 2023

Keywords

Cell Biology
Molecular Biology
Neuroscience

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

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10.1016/j.xpro.2023.102271

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

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",

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doi = "10.1016/j.xpro.2023.102271",

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T1 - Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody

AU - O'Leary, Justin

AU - Raulin, Ana Caroline

AU - Li, Zonghua

AU - Martens, Yuka

AU - Inoue, Yasuteru

AU - Strickland, Michael R.

AU - Han, Xianlin

AU - Holtzman, David M.

AU - Bu, Guojun

AU - Zhao, Na

PY - 2023/6/16

Y1 - 2023/6/16

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AB - 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.

KW - Cell Biology

KW - Molecular Biology

KW - Neuroscience

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

Abstract

Keywords

ASJC Scopus subject areas

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