Sensitive intranuclear flow cytometric quantification of IRF4 protein in multiple myeloma and normal human hematopoietic cells

STAR Protoc. 2021 May 27;2(2):100565. doi: 10.1016/j.xpro.2021.100565. eCollection 2021 Jun 18.

Abstract

Interferon regulatory factor 4 (IRF4) is a transcription factor that regulates normal and malignant immune cell development and is implicated in multiple myeloma pathogenesis. This protocol describes the use of combined cell surface and intranuclear staining with fluorescent antibodies to measure IRF4 protein expression within myeloma and normal immune cells. IRF4 protein quantification may provide a valuable prognostic tool to predict disease severity and sensitivity to IRF4-targeted therapies. This flow-cytometry-based procedure could also be rapidly translated into a clinically compatible assay. For complete details on the use and execution of this protocol, please refer to Mondala et al. (2021).

Keywords: Cancer; Cell isolation; Flow Cytometry/Mass Cytometry; Immunology; Stem Cells.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Marrow Cells / metabolism*
  • Flow Cytometry / methods*
  • Humans
  • Interferon Regulatory Factors / metabolism*
  • Limit of Detection
  • Multiple Myeloma / metabolism*
  • Multiple Myeloma / pathology

Substances

  • Interferon Regulatory Factors
  • interferon regulatory factor-4