Separation technology of proteins from a complex mixture by two-dimensional gel (2D gel) was invented more than 40 years ago. With a good laboratory practice, the 2D gels are likely to be dried and stored at ambient temperature as archived record. Up until the beginning of this century, it had been difficult to identify the protein spots isolated on 2D gels. However, the advent of mass spectrometry-based proteomics protocols combined with genome information enabled us to determine the identity of a protein separated on 2D gels archived decades ago. The protocol will assist researchers to decipher molecular mechanisms involved in the system by identifying and quantifying the protein of interest from archived 2D gels.
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