Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins

Molecular-level understanding of plasma cell (PC) differentiation has been modeled using lipopolysaccharide (LPS) stimulation in vitro. However, this system does not involve the B-cell receptor (BCR)—a critical component of B cell biology. Here, we present a protocol for in vitro PC differentiation...

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Autores principales: Ochiai, Kyoko, Shima, Hiroki, Ikura, Tsuyoshi, Franke, Marissa C., Sievert, Evelyn P., Sciammas, Roger, Igarashi, Kazuhiko
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255920/
https://www.ncbi.nlm.nih.gov/pubmed/34258594
http://dx.doi.org/10.1016/j.xpro.2021.100633
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author Ochiai, Kyoko
Shima, Hiroki
Ikura, Tsuyoshi
Franke, Marissa C.
Sievert, Evelyn P.
Sciammas, Roger
Igarashi, Kazuhiko
author_facet Ochiai, Kyoko
Shima, Hiroki
Ikura, Tsuyoshi
Franke, Marissa C.
Sievert, Evelyn P.
Sciammas, Roger
Igarashi, Kazuhiko
author_sort Ochiai, Kyoko
collection PubMed
description Molecular-level understanding of plasma cell (PC) differentiation has been modeled using lipopolysaccharide (LPS) stimulation in vitro. However, this system does not involve the B-cell receptor (BCR)—a critical component of B cell biology. Here, we present a protocol for in vitro PC differentiation system dependent on BCR signaling that easily scales up for cell number-demanding applications, including protein complex purification. We describe how to set up this system and detail applications for endogenous complex purification of chromatin-associated proteins. For further details on the use and execution of this protocol, please refer to Sciammas et al. (2011) and Ochiai et al. (2018, 2020).
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spelling pubmed-82559202021-07-12 Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins Ochiai, Kyoko Shima, Hiroki Ikura, Tsuyoshi Franke, Marissa C. Sievert, Evelyn P. Sciammas, Roger Igarashi, Kazuhiko STAR Protoc Protocol Molecular-level understanding of plasma cell (PC) differentiation has been modeled using lipopolysaccharide (LPS) stimulation in vitro. However, this system does not involve the B-cell receptor (BCR)—a critical component of B cell biology. Here, we present a protocol for in vitro PC differentiation system dependent on BCR signaling that easily scales up for cell number-demanding applications, including protein complex purification. We describe how to set up this system and detail applications for endogenous complex purification of chromatin-associated proteins. For further details on the use and execution of this protocol, please refer to Sciammas et al. (2011) and Ochiai et al. (2018, 2020). Elsevier 2021-06-30 /pmc/articles/PMC8255920/ /pubmed/34258594 http://dx.doi.org/10.1016/j.xpro.2021.100633 Text en © 2021 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
Ochiai, Kyoko
Shima, Hiroki
Ikura, Tsuyoshi
Franke, Marissa C.
Sievert, Evelyn P.
Sciammas, Roger
Igarashi, Kazuhiko
Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins
title Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins
title_full Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins
title_fullStr Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins
title_full_unstemmed Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins
title_short Protocol for in vitro BCR-mediated plasma cell differentiation and purification of chromatin-associated proteins
title_sort protocol for in vitro bcr-mediated plasma cell differentiation and purification of chromatin-associated proteins
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8255920/
https://www.ncbi.nlm.nih.gov/pubmed/34258594
http://dx.doi.org/10.1016/j.xpro.2021.100633
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