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Fractionation of Proteins by Immobilized Metal Affinity Chromatography

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2D PAGE: Sample Preparation and Fractionation

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 424))

Summary

It is widely known that the progress of proteomics mostly depends on the development of more advanced and sensitive protein separation technologies. Immobilized metal affinity chromatography (IMAC) is a useful protein fractionation method used to enrich metal associated proteins. IMAC represents an affinity separation approach based on the interaction between proteins and metal ions immobilized on a solid support. By changing various metal ions and other experimental conditions such as pH and elution composition, IMAC can selectively isolate metal-binding protein fractions for further specific proteomic analysis. The combination of IMAC with other protein analytical technologies has been successfully applied in characterizing metalloproteome and post-translational modifications. In the future, newly developed IMAC techniques integrated with other proteomic methods will significantly contribute to the further development of proteomics.

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References

  1. Sun,X., Chiu,J.F., and He,Q.Y. (2005) Application of immobilized metal affinity chromatography in proteomics. Expert Rev.Proteomics., 2, 649–657.

    Article  CAS  PubMed  Google Scholar 

  2. Posewitz,M.C. and Tempst,P. (1999) Immobilized gallium(III) affinity chromatography of phosphopeptides. Analytical Chemistry, 71, 2883–2892.

    Article  CAS  PubMed  Google Scholar 

  3. Scanff,P., Yvon,M., and Pelissier,J.P. (1991) Immobilized Fe3+ Affinity Chromatographic Isolation of Phosphopeptides. Journal of Chromatography, 539, 425–432.

    Article  CAS  PubMed  Google Scholar 

  4. Neville,D.C.A., Rozanas,C.R., Price,E.M., Gruis,D.B., Verkman,A.S., and Townsend,R.R. (1997) Evidence for phosphorylation of serine 753 in CFTR using a novel metal-ion affinity resin and matrix-assisted laser desorption mass spectrometry. Protein Science, 6, 2436–2445.

    Article  CAS  PubMed  Google Scholar 

  5. Ueda,E.K., Gout,P.W., and Morganti,L. (2003) Current and prospective applications of metal ion-protein binding. J Chromatogr.A, 988, 1–23.

    Article  CAS  PubMed  Google Scholar 

  6. Gaberc-Porekar,V. and Menart,V. (2001) Perspectives of immobilized-metal affinity chromatography. J.Biochem.Biophys.Methods, 49, 335–360.

    Article  CAS  PubMed  Google Scholar 

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© 2008 Humana Press, a part of Springer Science+Business Media, LLC

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Sun, X., Chiu, JF., He, QY. (2008). Fractionation of Proteins by Immobilized Metal Affinity Chromatography. In: Posch, A. (eds) 2D PAGE: Sample Preparation and Fractionation. Methods in Molecular Biology™, vol 424. Humana Press. https://doi.org/10.1007/978-1-60327-064-9_17

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  • DOI: https://doi.org/10.1007/978-1-60327-064-9_17

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-722-8

  • Online ISBN: 978-1-60327-064-9

  • eBook Packages: Springer Protocols

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