This fully functional N-terminal FLAG-tagged ubiquitin protein allows for the convenient detection or affinity purification of ubiquitinated proteins in vitro. The epitope tag is a hydrophilic octapeptide (DYKDDDDK) derived from the sequence of the bacteriophage 7 gene-10 product. This tag is specifically recognized by anti-FLAG antibodies M1, M2 or M5. Ubiquitin is a 76 amino acid, highly conserved nuclear and cytoplasmic protein. It is found exclusively in eukaryotes, becomes covalently attached to substrate proteins by enzymes in the Ubiquitin-Proteosome Pathway (UPP) and has a major role in targeting cellular proteins for the ATP-dependent degradation by the 26S proteosome. Ubiquitination also affects proteosomeindependent events such as protein localization, activity and function.