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NanoBRET™ Proteasomal Recruitment Assay Technical Manual

Instructions for Use of Product(s)

Literature # TM612

Most eukaryotic proteins are degraded by recruiting and processing mediated by the 26S proteasome. The NanoBRET™ Proteasomal Recruitment Assay is a live-cell assay that can monitor target proteins recruited to the 26S proteasome. This assay can also be used to study dynamic changes in 26S trafficking after cellular treatments, such as small molecules or pathway inducers, that would influence protein stabilization or degradation respectively.

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 of the dynamics of protein stabilization or degradation, the optimal NanoBRET™ assay configuration is with the target protein as the luminescent donor and the 26S proteasomal component as the fluorescent acceptor. This means you can monitor target protein levels while simultaneously observing proteasomal recruitment, made possible due to the luminescent to fluorescent ratio in the NanoBRET™ assay.

Printed in USA 3/20.