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

Harkness, Linda, Prokhorova, Tatyana A., Kassem, Moustapha and Blagoev, Blagoy (2012). Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis. In Kursad Turksen (Ed.), Human embryonic stem cells handbook (pp. 297-305) New York, NY, United States: Springer. doi:10.1007/978-1-61779-794-1_20


Author Harkness, Linda
Prokhorova, Tatyana A.
Kassem, Moustapha
Blagoev, Blagoy
Title of chapter Stable isotope labelling with amino acids in cell culture for human embryonic stem cell proteomic analysis
Title of book Human embryonic stem cells handbook
Place of Publication New York, NY, United States
Publisher Springer
Publication Year 2012
Sub-type Research book chapter (original research)
DOI 10.1007/978-1-61779-794-1_20
Open Access Status Not Open Access
Series Methods in Molecular Biology
ISBN 9781617797934
9781617797941
ISSN 1064-3745
Editor Kursad Turksen
Volume number 873
Chapter number 20
Start page 297
End page 305
Total pages 9
Total chapters 20
Subjects 1312 Molecular Biology
1311 Genetics
Abstract/Summary 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.
Keyword Human embryonic stem cells
Mass spectroscopy
Proteomics
Stable isotope labelling
Q-Index Code B1
Q-Index Status Provisional Code
Institutional Status Non-UQ

 
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Created: Thu, 24 Sep 2015, 01:07:21 EST by Linda Harkness on behalf of Examinations