Relative Quantification Mass Spectrometry Using iTRAQ Isobaric Tags View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2012-06-08

AUTHORS

Richard D. Unwin , Emma Carrick , Anthony D. Whetton

ABSTRACT

Advances in mass spectrometry, separation technologies and availability of genome sequences for a number of species now offer opportunities for in depth analysis of proteomes. The sensitivity of new instrumentation means detection of low copy number proteins such as transcription factors is well developed. As biology and biomedicine are quantitative disciplines there remains the question of methods for relative or absolute quantification of these proteins. Mass spectrometry per se is not amenable to relative quantification within complex protein mixtures meaning novel approaches are required. Methods for relative quantification such as spectral counting are currently being investigated but chemical labelling methods such as isobaric tagging offer a more robust workflow allowing relative quantification of multiple samples in a single tandem mass spectrometry run. Here we describe an approach, isobaric tagging for relative and absolute quantification (iTRAQ), that has been applied with success to hematopoietic stem cell and embryonic stem cell research, as well as in clinical proteomics studies and has been used to highlight the role for post-translational regulation of protein levels in stem cell development. In this chapter, we will outline the principle of the iTRAQ methodology, provide examples of its use and discuss it strengths and weaknesses. More... »

PAGES

77-95

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/978-94-007-4330-4_5

DOI

http://dx.doi.org/10.1007/978-94-007-4330-4_5

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1034727117


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