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

Publication Title

STAR Protoc

Authors

Mark A Gillespie, Institute for Systems Biology, Seattle, WA, USAFollow
Carmen G Palii
Daniel Sanchez-Taltavull
Theodore J Perkins
Marjorie Brand
Jeffrey A Ranish, Institute for Systems Biology, Seattle, WA 98109, USA.Follow

Document Type

Article

Publication Date

12-18-2020

Keywords

washington; seattle; ISB

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).

Specialty/Research Institute

Institute for Systems Biology