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

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 apo...

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Detalles Bibliográficos
Autores principales: O’Leary, Justin, Raulin, Ana-Caroline, Li, Zonghua, Martens, Yuka, Inoue, Yasuteru, Strickland, Michael R., Han, Xianlin, Holtzman, David M., Bu, Guojun, Zhao, Na
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10323120/
https://www.ncbi.nlm.nih.gov/pubmed/37289593
http://dx.doi.org/10.1016/j.xpro.2023.102271
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author O’Leary, Justin
Raulin, Ana-Caroline
Li, Zonghua
Martens, Yuka
Inoue, Yasuteru
Strickland, Michael R.
Han, Xianlin
Holtzman, David M.
Bu, Guojun
Zhao, Na
author_facet O’Leary, Justin
Raulin, Ana-Caroline
Li, Zonghua
Martens, Yuka
Inoue, Yasuteru
Strickland, Michael R.
Han, Xianlin
Holtzman, David M.
Bu, Guojun
Zhao, Na
author_sort O’Leary, Justin
collection PubMed
description 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.
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spelling pubmed-103231202023-07-07 Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody O’Leary, Justin Raulin, Ana-Caroline Li, Zonghua Martens, Yuka Inoue, Yasuteru Strickland, Michael R. Han, Xianlin Holtzman, David M. Bu, Guojun Zhao, Na STAR Protoc Protocol 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. Elsevier 2023-06-07 /pmc/articles/PMC10323120/ /pubmed/37289593 http://dx.doi.org/10.1016/j.xpro.2023.102271 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
O’Leary, Justin
Raulin, Ana-Caroline
Li, Zonghua
Martens, Yuka
Inoue, Yasuteru
Strickland, Michael R.
Han, Xianlin
Holtzman, David M.
Bu, Guojun
Zhao, Na
Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody
title Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody
title_full Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody
title_fullStr Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody
title_full_unstemmed Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody
title_short Standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein E particles utilizing HJ15.4 monoclonal antibody
title_sort standardized immunoprecipitation protocol for efficient isolation of native apolipoprotein e particles utilizing hj15.4 monoclonal antibody
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10323120/
https://www.ncbi.nlm.nih.gov/pubmed/37289593
http://dx.doi.org/10.1016/j.xpro.2023.102271
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