Thanks to its capability to recruit the natural protein degradation system of a cell, PROTACs (PROteolysis TArgeting Chimera) appears as a very promising technology in cancer therapy and disease treatment. As a distributor of innovative reagents for life science researchers, tebu-bio provides you with all the necessary tools to develop or test your own PROTACs system.
Detecting ubiquitylation
Ubitest Flow chart – LifeSensors at tebu-bio.com
Determining the linkage of polyubiquitin on target proteins is challenging. The traditional methods are either through Mass Spectrometry or immunoblot with linkage specific antibodies, which are cumbersome. LifeSensors has developed the UbiTest assay, a more definitive method for demonstrating the ubiquitylation linkage of a protein, which is to couple immunoprecipitation of polyubiquitylated protein with digestion by a linkage specific deubiquitylase prior to immunoblot analysis. An increased signal for the unmodified substrate or a decreased signal of polyubiquitylated substrate at high molecular weight after K48/K63 specific DUB treatment is a clear indication that the protein was K48/K63 ubiquitylated.