The Absolute Quantification Strategy: Application to Phosphorylation Profiling of Human Separase Serine 1126
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Last Update: 2021-01-03
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Source: Internet
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Author: User
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The absolute quantification (AQUA) strategy provides a means to determine the precise protein or modified protein levels directlyfrom cells or tissues. The technique is based on two major principles: stable isotope dilution theory and the use of syntheticpeptides containing such stable isotopes to exactly mimic native counterparts after proteolysis. These peptides can be synthesizedwith modifications such as phosphorylation, methylation, and acetylation to allow for the direct, quantitative analysis ofposttranslationally modified proteins. In this chapter, we discuss the development of an AQUA method and demonstrate its usefulnessin the measurement of endogenous levels of the human protein separase at a functionally relevant phosphorylation site, serine1126.
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