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Two-Dimensional Electrophoresis and Mass Spectrometry for Protein Identification

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Progenitor Cells

Part of the book series: Methods in Molecular Biology ((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.

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References

  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 1969/07/01

    CAS  PubMed  Google Scholar 

  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 1969/03/15

    Article  CAS  Google Scholar 

  3. Mets LJ, Bogorad L (1974) Two-dimensional polyacrylamide gel electrophoresis: an improved method for ribosomal proteins. Anal Biochem 57(1):200–210; Epub 1974/01/01

    Article  CAS  Google Scholar 

  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 1973/05/01

    Article  CAS  Google Scholar 

  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 1970/08/01

    Article  CAS  Google Scholar 

  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 1974/01/03

    Article  CAS  Google Scholar 

  7. O’Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250(10):4007–4021; Epub 1975/05/25

    PubMed  PubMed Central  Google Scholar 

  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 1982/09/01

    Article  CAS  Google Scholar 

  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 1985/09/01

    CAS  PubMed  PubMed Central  Google Scholar 

  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 1988/09/01

    Article  CAS  PubMed  Google Scholar 

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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.

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Authors and Affiliations

  1. Chicago College of Pharmacy, Midwestern University, Downers Grove, IL, USA

    Amaresh K. Ranjan & Anil Gulati

Authors
  1. Amaresh K. Ranjan
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  2. Anil Gulati
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Corresponding author

Correspondence to Amaresh K. Ranjan .

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Editors and Affiliations

  1. NHMRC Clinical Trials Centre, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW, Australia

    Mugdha V. Joglekar

  2. NHMRC Clinical Trials Centre, Faculty of Medicine and Health, The University of Sydney, Camperdown, NSW, Australia

    Anandwardhan A. Hardikar

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Ranjan, A.K., Gulati, A. (2019). Two-Dimensional Electrophoresis and Mass Spectrometry for Protein Identification. In: Joglekar, M., Hardikar, A. (eds) Progenitor Cells. Methods in Molecular Biology, vol 2029. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9631-5_15

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  • DOI: https://doi.org/10.1007/978-1-4939-9631-5_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9630-8

  • Online ISBN: 978-1-4939-9631-5

  • eBook Packages: Springer Protocols

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