What is MudPIT?
Multidimensional protein identification technology. Also referred as 2D-LC-MS. In MudPIT, a complex protein mixture is digested with a specific protease (most commonly Trypsin). The resulting peptide fragments then undergo two-dimensional liquid chromatography, which separates the peptides initially by charge, then by hydrophobicity to allow adequate separation. The column elutes into an Electrospray mass spectroscopy unit that identifies peptides as they elute, one by one, based on their tandem mass spectroscopy spectrum. Databases of peptides can then be searched to identify the proteins based on the mass spectra of the peptides. This technique is very sensitive and reproducible(in the femtomolar, 10-15 range, with a dynamic range of 104 and CV of 0.5%).