Abstract
Knowledge of the differences between the amounts and types of protein that are expressed in diseased compared to healthy subjects may give an understanding of the biological pathways that cause disease. This is the reasoning behind the presented protocol, which uses difference gel electrophoresis (DIGE) to discover up- or down-regulated proteins between mice of different genotypes, or of those fed on different diets, that may thus be prone to develop diabetes-like phenotypes. Subsequent analysis of these proteins by tandem mass spectrometry typically facilitates their identification with a high degree of confidence.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wild, S., Roglic, G., Green, A., Sicree, R., and King, H. (2004) Global prevalence of diabetes: estimates for the year 2000 and projections for 2030, Diabetes Care 27, 1047–1053.
WHO. (2009) Fact Sheet No. 312: Diabetes.
Bradford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal Biochem 72, 248–254.
Candiano, G., Bruschi, M., Musante, L., Santucci, L., Ghiggeri, G. M., Carnemolla, B., Orecchia, P., Zardi, L., and Righetti, P. G. (2004) Blue silver: a very sensitive colloidal Coomassie G-250 staining for proteome analysis, Electro-phoresis 25, 1327–1333.
Bindschedler, L. V., Palmblad, M., and Cramer, R. (2008) Hydroponic isotope labelling of entire plants (HILEP) for quantitative plant proteomics; an oxidative stress case study, Phytochemistry 69, 1962–1972.
Acknowledgements
This work was supported by the EU 6th Framework Programme “System-wide analysis and modelling of protein modification”. The authors would additionally like to thank colleagues at the University of Reading (The BioCentre, Department of Chemistry and ICMR) and former colleagues at University College London (Proteomics Unit) for their invaluable advice.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Smith, C., Mills, D., Cramer, R. (2012). Application of DIGE and Mass Spectrometry in the Study of Type 2 Diabetes Mellitus Mouse Models. In: Cramer, R., Westermeier, R. (eds) Difference Gel Electrophoresis (DIGE). Methods in Molecular Biology, vol 854. Humana Press. https://doi.org/10.1007/978-1-61779-573-2_21
Download citation
DOI: https://doi.org/10.1007/978-1-61779-573-2_21
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-572-5
Online ISBN: 978-1-61779-573-2
eBook Packages: Springer Protocols