Based on protein domains known to possess an affinity for ubiquitin, Tandem Ubiquitin Binding Entities (TUBEs) have been developed for the isolation and identification of ubiquitylated proteins. TUBEs display up to a 1000-fold increase in affinity for poly-ubiquitin moieties over the single ubiquitin binding associated domain (UBA). In addition, TUBEs display a protective effect on polyubiquitinated proteins, allowing for detection at relatively low abundance. These properties effectively “capture” proteins in their polyubiquitylated state. TUBE 2 is based on UBA1 from the human RAD23A. We have developed the HRP-TUBE 2 for quick and easy detection of polyubiquitin and polyubiquitinated substrates.
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