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

Kyoko Ochiai; Hiroki Shima; Tsuyoshi Ikura; Marissa C. Franke; Evelyn P. Sievert; Roger Sciammas; Kazuhiko Igarashi

doi:10.1016/j.xpro.2021.100633

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

Kyoko Ochiai, Hiroki Shima, Tsuyoshi Ikura, Marissa C. Franke, Evelyn P. Sievert, Roger Sciammas, Kazuhiko Igarashi

MED - Medical Sciences

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

1 Citation (Scopus)

Abstract

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).

Original language	English
Pages (from-to)	100633
Number of pages	1
Journal	STAR Protocols
Volume	2
Issue number	3
DOIs	https://doi.org/10.1016/j.xpro.2021.100633
Publication status	Published - 2021 Sept 17

Keywords

Antibody
Cell Biology
Cell Differentiation
Chromatin immunoprecipitation (ChIP)
Flow Cytometry/Mass Cytometry
Gene Expression
Immunology
Mass Spectrometry
Molecular Biology

ASJC Scopus subject areas

Medicine(all)

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10.1016/j.xpro.2021.100633

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

abstract = "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).",

keywords = "Antibody, Cell Biology, Cell Differentiation, Chromatin immunoprecipitation (ChIP), Flow Cytometry/Mass Cytometry, Gene Expression, Immunology, Mass Spectrometry, Molecular Biology",

author = "Kyoko Ochiai and Hiroki Shima and Tsuyoshi Ikura and Franke, {Marissa C.} and Sievert, {Evelyn P.} and Roger Sciammas and Kazuhiko Igarashi",

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doi = "10.1016/j.xpro.2021.100633",

language = "English",

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AU - Ochiai, Kyoko

AU - Shima, Hiroki

AU - Ikura, Tsuyoshi

AU - Franke, Marissa C.

AU - Sievert, Evelyn P.

AU - Sciammas, Roger

AU - Igarashi, Kazuhiko

PY - 2021/9/17

Y1 - 2021/9/17

N2 - 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).

AB - 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).

KW - Antibody

KW - Cell Biology

KW - Cell Differentiation

KW - Chromatin immunoprecipitation (ChIP)

KW - Flow Cytometry/Mass Cytometry

KW - Gene Expression

KW - Immunology

KW - Mass Spectrometry

KW - Molecular Biology

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DO - 10.1016/j.xpro.2021.100633

M3 - Article

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SN - 2666-1667

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SP - 100633

JO - STAR Protocols

JF - STAR Protocols

IS - 3

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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