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

Research output: Contribution to journalArticlepeer-review

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 languageEnglish (US)
Pages (from-to)100633
Number of pages1
JournalSTAR Protocols
Volume2
Issue number3
DOIs
StatePublished - Sep 17 2021

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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