Sample Fractionation with ZOOM® IEF Fractionator

A fast, easy-to-use, and versatile method for isoelectric focusing of complex protein samples

The ZOOM® IEF Fractionator reduces highly complex protein samples into fractions, based upon isoelectric point, for analysis by two dimensional gel electrophoresis (2DE), one dimensional gel electrophoresis (1DE), or two dimensional liquid chromatography/mass spectrometry (2D LC/MS). The ZOOM® IEF Fractionator offers a versatile range of separation (fractionation) options, in as little as three hours.

Solution phase isoelectric focusing with the ZOOM® IEF Fractionator provides reproducible and cost-effective separations that:

  • Allow for loading of increased amounts of protein for downstream applications
  • Enrich low abundance proteins and increase the dynamic range of detection
  • Reduce precipitation/aggregation artifacts associated with high protein load samples

ZOOM® Disks are pre-cast polyacrylamide gels, eliminating the need for manual preparation and minimizing the chance of cross-contamination. These immobilized buffered disks are pre-labeled, disposable, and designed for single use, ensuring consistent and reproducible fractionation. Using seven ZOOM® Disks, of specific pH, protein samples can be resolved into six fractions from pH 3 – 12.

Integrating the IEF Fractionator into the 2D Workflow

Integrating IEF Fractionato

The ZOOM® Basic Protein Kit facilitates fractionation of proteins in the pH 9 – 12 range, resulting in concentrated protein fractions that are free of salts and other interfering proteins, for analysis by 2DE, 1DE, and by mass spectrometry following a precipitation procedure.

References

  1. Zuo, X. and Speicher, D.W. (2000) Anal. Biochem. 284: 266-278.
  2. Zuo, et. al. (2001) Electrophoresis 22: 1603-1615.
  3. Zuo, X. and Speicher, D.W. (2002) Proteomics: 2: 58-68.
  4. Zuo, X., Hembach, P., Echan, L., and Speicher, D.W. (2002) Journal of Chromatography B 782: 253-265.
  5. Ali-Khan, N., Zuo, X., and Speicher, D.W. (2002) Current Protocols in Protein Science 22.1: 1-19.
  6. Zuo, X. and Speicher, D.W. (2000) Methods in Molecular Biology (Humana Press, P. Cutler, ed.) In press.
  7. Zuo, X. and Speicher D.W. 2004. Microscale solution isoelectrofocusing: A sample prefractionation method for comprehensive proteome analysis. In: Methods in Molecular Biology: Protein Purification Protocols. Humana Press, Totowa, NJ (P. Cutler, ed.), pp.361-375.
  8. Speicher, D.W. 2004. Overview of proteome analysis. In: Proteome Analysis: Interpreting the Genome (D. Speicher, ed.). Elsevier, The Netherlands, pp. 1-18.
  9. Zuo, X., Lee, K., Speicher, D.W. 2004. Electrophoretic prefractionation for comprehensive analysis of proteomes. In: Proteome Analysis: Interpreting the Genome (D. Speicher, ed.). Elsevier, The Netherlands, pp. 93-118.
  10. Speicher, D.W., Lee, K., Tang, H.-Y., Echan, L., Ali-Khan, N., Zuo, X., and Hembach, P. 2004. Current challenges in proteomics: Mining low abundance proteins and expanding protein profiling capacities. In: Advances in Mass Spectrometry, Vol. 16 (Ashcroft, A.E., Brenton, G, and Monaghan, J.J., ed.) pp. 37-57.
  11. Zuo, X., Lee, K., and Speicher, D.W. 2004. Fractionation of complex proteomes by microscale solution isoelectrofocusing using ZOOM® IEF fractionators to improve protein profiling. In: The Proteome Handbook (Walker, J.M., ed.) In press.