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Two-Dimensional Electrophoresis and Mass Spectrometry for Protein Identification

  • Amaresh K. RanjanEmail author
  • Anil Gulati
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2029)

Abstract

Two-dimensional electrophoresis is useful for separation of individual proteins that can be easily isolated and processed with mass spectrometry for their identification. Here we describe a simplified method of 2D electrophoresis with aim to help beginners in optimization of protein separation protocol, which can be further modified to enhance protein spot resolution. Current protocol can be used in different fields of biology including progenitor cells.

Key words

2D Electrophoresis Proteins Proteomics Mass spectrometry Isoelectric focusing SDS-PAGE 

Notes

Acknowledgments

We acknowledge Dipavo Banerjee and Divya Khandekar for manuscript preparation. We thank National Centre for Cell Science, Pune, India, for supporting the related original diabetes research.

References

  1. 1.
    Macko V, Stegemann H (1969) Mapping of potato proteins by combined electrofocusing and electrophoresis identification of varieties. Hoppe Seylers Z Physiol Chem 350(7):917–919; Epub 1969/07/01PubMedGoogle Scholar
  2. 2.
    Margolis J, Kenrick KG (1969) 2-dimensional resolution of plasma proteins by combination of polyacrylamide disc and gradient gel electrophoresis. Nature 221(5185):1056–1057; Epub 1969/03/15CrossRefGoogle Scholar
  3. 3.
    Mets LJ, Bogorad L (1974) Two-dimensional polyacrylamide gel electrophoresis: an improved method for ribosomal proteins. Anal Biochem 57(1):200–210; Epub 1974/01/01CrossRefGoogle Scholar
  4. 4.
    Orrick LR, Olson MO, Busch H (1973) Comparison of nucleolar proteins of normal rat liver and Novikoff hepatoma ascites cells by two-dimensional polyacrylamide gel electrophoresis. Proc Natl Acad Sci U S A 70(5):1316–1320; Epub 1973/05/01CrossRefGoogle Scholar
  5. 5.
    Kaltschmidt E, Wittmann HG (1970) Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins. Anal Biochem 36(2):401–412; Epub 1970/08/01CrossRefGoogle Scholar
  6. 6.
    MacGillivray AJ, Rickwood D (1974) The heterogeneity of mouse-chromatin nonhistone proteins as evidenced by two-dimensional polyacrylamide-gel electrophoresis and ion-exchange chromatography. Eur J Biochem 41(1):181–190; Epub 1974/01/03CrossRefGoogle Scholar
  7. 7.
    O’Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250(10):4007–4021; Epub 1975/05/25PubMedPubMedCentralGoogle Scholar
  8. 8.
    Bjellqvist B, Ek K, Righetti PG, Gianazza E, Gorg A, Westermeier R et al (1982) Isoelectric focusing in immobilized pH gradients: principle, methodology and some applications. J Biochem Biophys Methods 6(4):317–339; Epub 1982/09/01CrossRefGoogle Scholar
  9. 9.
    Gorg A, Postel W, Weser J, Patutschnick W, Cleve H (1985) Improved resolution of PI (alpha 1-antitrypsin) phenotypes by a large-scale immobilized pH gradient. Am J Hum Genet 37(5):922–930; Epub 1985/09/01PubMedPubMedCentralGoogle Scholar
  10. 10.
    Gorg A, Postel W, Gunther S (1988) The current state of two-dimensional electrophoresis with immobilized pH gradients. Electrophoresis 9(9):531–546.  https://doi.org/10.1002/elps.1150090913; Epub 1988/09/01CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Chicago College of PharmacyMidwestern UniversityDowners GroveUSA

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