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NanoBRET™ Ubiquitination Assay Technical Manual

Instructions for Use of Product(s)

Literature # TM616

The NanoBRET™ Ubiquitination Assay is a live-cell ubiquitin assay, broadly measuring all types of ubiquitination on a target protein, both in terms of extent (mono- and poly-ubiquitination) and amino acid linkage, but cannot discern amongst them. This assay can also be used to dynamically detect either an increase or a decrease in the relative level of target ubiquitination after cell treatments, including induction of signaling pathways or compound treatment.

This assay uses NanoBRET™ technology, a proximity-based method dependent upon energy transfer from a luminescent donor to a fluorescent acceptor that is measured using an instrument capable of reading dual-filtered luminescence. Because ubiquitination potentially leads to protein loss, the optimal NanoBRET™ Ubiquitination Assay configuration is with the target protein as the luminescent donor and the ubiquitin protein as the fluorescent acceptor. This means you can monitor potential protein loss of the target protein while simultaneously assessing ubiquitination, made possible due to luminescent to fluorescent ratio in the NanoBRET™ assay.

Printed in USA 3/20.