Abstract
The field of proteomics acts as a critical tool to unravel the vast range of proteomes to systematically investigate the dynamics of cells. The evolution of gel-based techniques, such as PAGE, SDS-PAGE, IEFs, and 2DGEs, is still employed in mainstream research fields for investigating the proteomes of living organisms. The advancements in these techniques led to offer a new window of gel-free techniques to overcome the limitations of gel-based techniques. The gel-free proteomic techniques such as isotope-coded affinity tags, isotope-coded protein labeling, isobaric tags for relative and absolute quantification, tandem mass tag, and stable isotopic labeling of amino acids in cell culture have characterized the proteomes more accurately and precisely to broaden the applicability of proteomic approaches. With this background, this chapter aimed to have the technical know-how of major gel-based and gel-free proteomic approaches for the characterization of biomolecules for investigating living systems on multiple levels. The technical advances in these techniques have led to understand the working manual of proteomics to understand the diversity of proteins existing in complex eukaryotic systems.
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