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

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
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

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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",

author = "Justin O'Leary and Raulin, \{Ana Caroline\} and Zonghua Li and Yuka Martens and Yasuteru Inoue and Strickland, \{Michael R.\} and Xianlin Han and Holtzman, \{David M.\} and Guojun Bu and Na Zhao",

note = "Publisher Copyright: {\textcopyright} 2023 The Author(s)",

year = "2023",

month = jun,

day = "16",

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

language = "English",

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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

N2 - 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.

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

UR - https://www.scopus.com/pages/publications/85161057715

U2 - 10.1016/j.xpro.2023.102271

DO - 10.1016/j.xpro.2023.102271

M3 - Article

C2 - 37289593

AN - SCOPUS:85161057715

SN - 2666-1667

VL - 4

JO - STAR Protocols

JF - STAR Protocols

IS - 2

M1 - 102271

ER -

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

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