Quantification of pluripotency transcription factor levels in embryonic stem cells by flow cytometry

Nicola Festuccia, Ian Chambers

Research output: Contribution to journalArticlepeer-review

Abstract

Embryonic stem (ES) cell lines are derived from the inner cell mass of the pre-implantation blastocyst and are characterized by the ability to undergo indefinite self-renewal while retaining the potential to differentiate into each of the three primary germ layers. The ability of individual ES cells to self-renew or appropriately respond to differentiation signals is influenced by the intracellular level of a number of crucial transcription factors. It is therefore important to be able to reliably quantify the levels of these proteins in single cells. Here we present an intracellular staining technique for flow cytometry suitable for monitoring transcription factor expression in ES cells. We illustrate the application of this technique to the detection of Oct4 and Nanog proteins and the coupling of this approach with fluorescent reporters of gene activity.
Original languageEnglish
Pages (from-to)1B.9.1-1B.9.13
JournalCurrent protocols in stem cell biology
Volume19
Issue number1
Early online date1 Dec 2011
DOIs
Publication statusPublished - 31 Dec 2011

Keywords

  • Octamer Transcription Factor-3
  • Pluripotent Stem Cells
  • Animals
  • Homeodomain Proteins
  • Transcription Factors
  • Humans
  • Cell Differentiation
  • Flow Cytometry
  • Embryonic Stem Cells
  • Staining and Labeling
  • Cell Line
  • ES cells
  • FACS
  • quantition
  • intracellular staining
  • Nanog Oct4

Fingerprint

Dive into the research topics of 'Quantification of pluripotency transcription factor levels in embryonic stem cells by flow cytometry'. Together they form a unique fingerprint.

Cite this