2-D electrophoresis
In proteomics, 2-D electrophoresis is used to separate proteins by their isoelectric point, and then by their mass. This procedure has two main applications: simultaneous identification of mass quantities of proteins, and Coomassie Blue or silver stained for viewing the protein spots. All gels are then scanned using a GS-800 calibrated densitometerfrom which one can obtain TIFF files of the image. We also can perform image analysis using PDQuest software version 7.0 from Bio-Rad. This software permits the comparison and analysis of replicates of gels to determine statistically and scientifically meaningful spots. Proteins of interest then can be excised from the gel, characterized and identified by mass spectrometry.
To summarize, 2-D gel electrophoresis process consists of these steps:
- Sample preparation
- First dimension isoelectric focusing
- Second dimension gel electrophoresis
- Staining
- Imaging analysis
See also: electrophoresis
