Absolute quantification of transcription factors in human erythropoiesis using selected reaction monitoring mass spectrometry

STAR Protoc. 2020 Dec 13;1(3):100216. doi: 10.1016/j.xpro.2020.100216. eCollection 2020 Dec 18.

Abstract

Quantitative changes in transcription factor (TF) abundance regulate dynamic cellular processes, including cell fate decisions. Protein copy number provides information about the relative stoichiometry of TFs that can be used to determine how quantitative changes in TF abundance influence gene regulatory networks. In this protocol, we describe a targeted selected reaction monitoring (SRM)-based mass-spectrometry method to systematically measure the absolute protein concentration of nuclear TFs as human hematopoietic stem and progenitor cells differentiate along the erythropoietic lineage. For complete details on the use and execution of this protocol, please refer to Gillespie et al. (2020).

Keywords: Cell Differentiation; Mass Spectrometry; Proteomics.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Differentiation / genetics
  • Erythropoiesis / physiology*
  • Gene Expression Regulation / genetics
  • Gene Regulatory Networks / genetics
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Mass Spectrometry / methods*
  • Proteomics / methods
  • Transcription Factors / analysis*

Substances

  • Transcription Factors