Abstract
Proteome analysis has been revolutionized by the marriage of two-dimensional (2-D) gel electrophoresis and mass spectrometry (1–4). Mass spectrometry in combination with database searching permits the large-scale identification of proteins, and 2-D gel electrophoresis allows for the large-scale visualization and separation of cellular proteins. Previously, spot quantitation has been difficult, and numbers obtained were usually comparative, but not absolute (5–7). Mass spectrometry and 2-D gel electrophoresis in tandem can be used to quantitate protein spots. This chapter covers protocols to quantitate precisely the relative amounts of protein observed in the 2-D gels using radiolabeled total cell extracts.
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© 1999 Humana Press Inc., Totowa, NJ
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Gygi, S.P., Aebersold, R. (1999). Absolute Quantitation of 2-D Protein Spots. In: Link, A.J. (eds) 2-D Proteome Analysis Protocols. Methods in Molecular Biology, vol 112. Humana Press. https://doi.org/10.1385/1-59259-584-7:417
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DOI: https://doi.org/10.1385/1-59259-584-7:417
Publisher Name: Humana Press
Print ISBN: 978-0-89603-524-9
Online ISBN: 978-1-59259-584-6
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