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Two Dimensional Gel Electrophoresis (2D)

2D Gel Images The analysis of a proteome begins from separating individual proteins, as well as gaining insights into their quantitative characteristics.  Two-Dimensional Gel Electrophoresis (2D) allows the separation of proteins based on pH by isoelectric focusing (IEF) on the first dimension, and based on size by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) on the second dimension.

During IEF, proteins are separated on an immobilized pH gradient (IPG) strip by migrating to their isoelectric point (pI), where their net charges are zero. IPG strips are available in different lengths to accommodate various gel sizes and different pH ranges to allow optimal target separation.

After IEF, the IPG strip will be transferred to a SDS-PAGE gel for the second dimensional protein separation by molecular sizes.  Different percentage of gel can be used for optimal separation of different target sizes.  The separated proteins are then visualized by various staining methods.

2D offers high resolution, superior reproducibility and the capacity for simultaneous separation of thousands of proteins on one gel.  It has been widely used for expression and quantitative analysis of protein mixtures.  It remains as the most common technique for proteomics research.