2-D DIGE:Best Practice in 2-D Electrophoresis

ITQB Seminar

Title: 2-D DIGE^TM: Best Practice in 2-D Electrophoresis

Speaker: Bruno Bacher

Affiliation: GE Healthcare Europe GmbH

Host: Carla Pinheiro- Senior Researcher at Plant Biochemistry Laboratory

Abstract:

Two-dimensional (2-DE) gel electrophoresis is a powerful technique which allows separation and visualization of thousands of proteins in a single gel . For many years, high system variability and limited sensitivity of classical 2-D gels made it challenging to measure expression changes quantitatively and 2-DE was rather used as a qualitative tool.

2-D DIGE is multiplexing 2-DE with DIGE CyDyesTM  for fluorescent protein labeling which allows to introduce an internal standard  to compensate gel-to-gel variation for each individual spot.  This standardization using the internal standard is key for a very accurate and sensitive detection of even small expression changes with high statistical confidence.

Here, the experimental workflow of 2-D DIGE is discussed as well as modern software tools for in depth data analysis.  Strategies for assessment of data quality are given together with practical examples adapting and optimizing the experimental design.

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OPEN - Training  Course in Differential Gel Electrophoresis       

http://www.itqb.unl.pt/news/training-course-in-differential-gel-electrophoresis