Current Strategies for Quantitative Proteomics

摘要: Publisher Summary The chapter focuses on the current strategies for quantitative proteomics. establishment of an absolute quantitation requires use internal standards known concentration, impossible scenario considering discovery-driven nature proteomic studies and complexity samples being analyzed. There are two primary techniques to measure relative abundances proteins from distinct cell populations. These make tools in proteomics today: two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) mass spectrometry (MS). 2D-PAGE is relatively straightforward. Protein extracts populations separated different gels. Two-dimensional important separation technique research, whereby qualitative, as well protein expressions can be investigated. broad identification capabilities these solution-based combined with MS have led development methods quantitate detected. most popular example methodologies utilizes a reagent referred “isotope-coded affinity tags” (ICAT) that allows global measurements peptide virtually any source. second relies proteins. This measurement commonly accomplished through stable isotopes differentially label sources. One exciting aspects more recent developments both gel-based labeling allow abundance changes measured systems. Gel-based technologies such difference (DIGE) melting profile (MP) enable thousands compared across systems precise high throughput manner.