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MAP Kinase Signaling Protocols pp 263–281Cite as

Methods in Functional Proteomics

Two-Dimensional Polyacrylamide Gel Electrophoresis With Immobilized pH Gradients, In-Gel Digestion, and Identification of Proteins by Mass Spectrometry

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 250))

Abstract

The recent combination of several technology platforms has revolutionized the analysis of expressed cellular proteins (proteomics). Large-scale separation and visualization of proteins is accomplished using two-dimensional gel electrophoresis (2-DE). Proteins are separated first by isoelectric focusing (IEF) using immobilized pH gradient (IPG) strips. Following charge separation, proteins are further resolved by molecular mass using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), as shown in Fig. 1.

2-DE gel pattern of WM35 (melanoma) cell line.

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Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO

    Karine R. Bernard, Karen R. Jonscher, Katheryn A. Resing & Natalie G. Ahn

  2. Howard Hughes Medical Institute, University of Colorado, Boulder, CO

    Natalie G. Ahn

Authors
  1. Karine R. Bernard
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  2. Karen R. Jonscher
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  3. Katheryn A. Resing
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  4. Natalie G. Ahn
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Editor information

Editors and Affiliations

  1. Department of Biological Regulation, The Zeizmann Institute of Science, Rehovot, Israel

    Rony Seger

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© 2004 Humana Press Inc., Totowa, NJ

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Bernard, K.R., Jonscher, K.R., Resing, K.A., Ahn, N.G. (2004). Methods in Functional Proteomics. In: Seger, R. (eds) MAP Kinase Signaling Protocols. Methods in Molecular Biology™, vol 250. Humana Press. https://doi.org/10.1385/1-59259-671-1:263

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  • DOI: https://doi.org/10.1385/1-59259-671-1:263

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-998-8

  • Online ISBN: 978-1-59259-671-3

  • eBook Packages: Springer Protocols

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