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Application of Selected Reaction Monitoring to Highly Multiplexed Targeted Quantitative Proteomics - A Replacement for Western Blot Analysis (Paperback, 2013 ed.)
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Application of Selected Reaction Monitoring to Highly Multiplexed Targeted Quantitative Proteomics - A Replacement for Western Blot Analysis (Paperback, 2013 ed.)
Series: SpringerBriefs in Systems Biology
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A key experiment in biomedical research is monitoring the
expression of different proteins in order to detect changes that
occur in biological systems under different experimental
conditions. The method that is most widely used is the Western blot
analysis. While Western blot is a workhorse in laboratories
studying protein expression and has several advantages, it also has
a number of significant limitations. In particular, the method is
semi-quantitative with limited dynamic range. Western blot focuses
on a single protein per sample with only a small number of
representative samples analyzed in an experiment. New quantitative
tools have been needed for some time to at least supplement, &
possibly replace, the Western blot. Mass spectrometric methods have
begun to compete with Western blot for routine quantitative
analyses of proteins. One of these methods is based on the tandem
mass spectrometry technique of selected reaction monitoring (SRM),
which is also called multiple reaction monitoring (MRM). Selected
reaction monitoring is actually an older tandem mass spectrometry
technique, first described in the late 70s, that is widely utilized
in the quantitative analysis of small molecules like drugs &
metabolites. The use of selected reaction monitoring for the
quantitative analysis of proteins has a number of advantages. Most
importantly, it is fundamentally quantitative with a wide dynamic
range. The output of the analysis is a numerical result that can
range over several orders of magnitude. Other advantages include
sufficient specificity & sensitivity to detect low abundance
proteins in complex mixtures. Finally, selected reaction monitoring
can be multiplexed to allow the quantitative analysis of relatively
large numbers of proteins in a single sample in a single
experiment. This Brief will explain both the theoretical &
experimental details of the selected reaction monitoring experiment
as it is applied to proteins.
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