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

Producción científica: Article › revisión exhaustiva

Resumen

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.

Idioma original	English (US)
Número de artículo	102271
Publicación	STAR Protocols
Volumen	4
N.º	2
DOI	https://doi.org/10.1016/j.xpro.2023.102271
Estado	Published - jun 16 2023

ASJC Scopus subject areas

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

Acceder al documento

10.1016/j.xpro.2023.102271

Otros archivos y enlaces

Citar esto

@article{14831e93454e49dc8ddeb25d6d4fc8d1,

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 (US)",

volume = "4",

journal = "STAR Protocols",

issn = "2666-1667",

publisher = "Cell Press",

number = "2",

}

TY - JOUR

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 - http://www.scopus.com/inward/record.url?scp=85161057715&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85161057715&partnerID=8YFLogxK

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

Resumen

ASJC Scopus subject areas

Acceder al documento

Otros archivos y enlaces

Huella

Citar esto