Why does using a pooled internal standard increase the precision of the technology?
With any biological technique the precision is greatly improved if an internal standard is used. This is one of the key benefits of 2-D DIGE, the only 2-D electrophoresis technique that allows the user to run an internal standard. When using a pooled internal standard, every protein in the population appears on each gel. Each experimental sample can then be compared internally to the same standard, thereby accounting for any gel-to-gel (system) variation. This facilitates fully automated data processing and accurate spot statistics when using DeCyder Differential Analysis Software. The inclusion of an internal standard on each gel also aids matching between gels. Occasionally, protein expression changes between samples are so large that the expression of specific proteins can be switched off. The use of the pooled internal standard with spot co-detection of multiplexed samples avoids any problems caused by presence-absence events. The pooled internal standard would always contain every
