Image pretreatment tools II: Normalization techniques for 2-DE and 2-D DIGE

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

Gel electrophoresis is usually applied to identify different protein expression profiles in biological samples (e.g., control vs. pathological, control vs. treated). Information about the effect to be investigated (a pathology, a drug, a ripening effect, etc.) is however generally confounded with experimental variability that is quite large in 2-DE and may arise fromsmall variations in the sample preparation, reagents, sample loading, electrophoretic conditions, staining and image acquisition. Obtaining valid quantitative estimates of protein abundances in each map, before the differential analysis, is therefore fundamental to provide robust candidate biomarkers. Normalization procedures are applied to reduce experimental noise and make the images comparable, improving the accuracy of differential analysis. Certainly, they may deeply influence the final results, and to this respect they have to be applied with care. Here, the most widespread normalization procedures are described both for what regards the applications to 2-DE and 2D Difference Gel-electrophoresis (2-D DIGE) maps.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages91-107
Number of pages17
DOIs
Publication statusPublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1384
ISSN (Print)1064-3745

Keywords

  • 2-D DIGE
  • Gel-electrophoresis
  • Normalization

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