As part of the Signal-Seeker™ product line, Ubiquitin affinity beads have been optimized in order to detect endogenous levels of mono- and poly-ubiqutinated proteins, which often represent <1% of the target protein. UBA01-Beads can enrich all ubiquitinated species with a high affinity, including mono-, multi- and polyubiquitin chains (see Application 1 below). The affinity moeities have been cross-linked to beads for low leaching and cleaner detection of the protein of interest. A comprehensive Signal-Seeker™ Ubiquitination Detection Kit is also available (BK161) and is recommended for first time users.
Figure 1: Swiss 3T3 cells were pre-treated with MG-132, then either untreated or treated with CNF1 (CN04) for 3 hours prior to lysis with BlastR buffer. The BK161 kit was utilized to perform the IP on 300 μg of lysate per condition. Lane 1: 3 μg input untreated lysate, Lane 2: Ubiquitin Affinity beads (UBA01) plus untreated lysate, Lane 3: UBA01 plus CN04 treated lysate, Lane 4: ubiquitin control beads (CUB02) plus untreated lysate, Lane 5: CUB02 plus CN04 treated lysate. Samples were analyzed for Rac1 ubiquitination using an anti-Rac1 antibody.
Each package contains enough ubiquitin beads for 40 reactions and sufficient control beads for 5 reactions.
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Signal-Seeker™ Ubiquitination Detection Kit (Cat. # BK161)
Signal-Seeker™ SUMOylation 2/3 Detection Kit (Cat. # BK162)
Signal-Seeker™ Phosphotyrosine Detection Kit (Cat. # BK160)
Signal-Seeker™: BlastR™ Rapid Lysate Prep Kit (Cat. # BLR01)
Signal-Seeker™: PTMtrue™ Ubiquitin Antibody (Cat.# AUB01)