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

STAR Protoc. 2021 Jun 30;2(3):100633. doi: 10.1016/j.xpro.2021.100633. eCollection 2021 Sep 17.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation*
  • Chromatin / metabolism*
  • Chromatography, Liquid / methods
  • Culture Media
  • Mice
  • Mice, Transgenic
  • Plasma Cells / cytology*
  • Proteins / isolation & purification*
  • Proteins / metabolism
  • Receptors, Antigen, B-Cell / genetics
  • Receptors, Antigen, B-Cell / metabolism*
  • Tandem Mass Spectrometry / methods

Substances

  • Chromatin
  • Culture Media
  • Proteins
  • Receptors, Antigen, B-Cell