Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis

Publikation: Kapitel i bog/rapport/konference-proceedingKapitel i bogForskning

Abstract

The identification and quantitative measurements of proteins in human embryonic stem cells (hESC) is a fast growing interdisciplinary area with an enormous impact on understanding the biology of hESC and the mechanism controlling self-renewal and differentiation. Using a quantitative mass spectroscopic method of stable isotope labelling with amino acids during cell culture (SILAC), we are able to analyse differential expression of proteins from different cellular compartments and to identify intracellular signalling pathways involved in self-renewal and differentiation. In this chapter, we provide a detailed method for creating SILAC media suitable for use in hESC experiments, additionally we describe methods for the isolation of membrane fractions and cytosolic and nuclear/membrane fractions.
OriginalsprogEngelsk
TitelHuman Embryonic Stem Cells Handbook
RedaktørerK Turksen
Antal sider9
ForlagHumana Press
Publikationsdato2012
Sider297-305
ISBN (Trykt)978-1-61779-793-4
ISBN (Elektronisk)978-1-61779-794-1
DOI
StatusUdgivet - 2012
NavnMethods in Molecular Biology
Vol/bind873
ISSN1064-3745

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