Chapter title |
DIGE Analysis of Human Tissues
|
---|---|
Chapter number | 11 |
Book title |
Difference Gel Electrophoresis
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7268-5_11 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7267-8, 978-1-4939-7268-5
|
Authors |
Cecilia Gelfi, Daniele Capitanio |
Abstract |
Two-dimensional difference gel electrophoresis (2-D DIGE) is an advanced and elegant gel electrophoretic analytical tool for comparative protein assessment. It is based on two-dimensional gel electrophoresis (2-DE) separation of fluorescently labeled protein extracts. The tagging procedures are designed to not interfere with the chemical properties of proteins with respect to their pI and electrophoretic mobility, once a proper labeling protocol is followed. The two-dye or three-dye systems can be adopted and their choice depends on specific applications. Furthermore, the use of an internal pooled standard makes 2-D DIGE a highly accurate quantitative method enabling multiple protein samples to be separated on the same two-dimensional gel. The image matching and cross-gel statistical analysis generates robust quantitative results making data validation by independent technologies successful. |
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