Development of environmental proteomics depends on the separated and test techniques. There are three techniques in environmental proteomics, they are two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS), protein chip, bioinformatics techniques. These techniques are high-throughput, high speed and high sensitive.Two-dimensional gel electrophoresis (2DE) and mass spectrometry (MS) are the prevalent techniques for separating and testing of proteins. The 25-years-old, labor-intensive 2-DE techniques is used to resolve large numbers of proteins in a quantitative fashion as the only established method available to most laboratories. This core technology has been gradual progress during the past year. Currently, 2-DE is the only technique that can reveal hundreds or even thousands of proteins at a time, and is already yielding important findings across a wide range of applications. 2-DE couples IEF in the first dimension with SDS-PAGE in the second dimension, and can separate the complex mixtures of proteins according to isoelectric point (pI), molecular mass (Mr), solubility, and relative abundance. The current state of 2-DE use the immobilized pH gradient (IPG) as the first dimension, and the protocol described in 1988 is still valid today: IEF is performed in individual IPG gel stripe, and after equilibration with SDS buffer in the presence of urea, glycerol, and iodoacetamide, they are applied to a horizontal or vertical SDS gel.
展开▼
