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NanoBRET™ TE Intracellular E3 Ligase Assays, CRBN and VHL, Technical Manual

Instructions for Use of Product(s)
N2910, N2911, N2912, N2930, N2931, N2932

Literature # TM626

The NanoBRET™ TE Intracellular E3 Ligase Assay measures compound binding to an E3 ubiquitin ligase protein, such as cereblon (CRBN) or von Hippel-Lindau disease tumor suppressor (VHL), fused to NanoLuc® luciferase. The NanoBRET™ TE E3 Ligase Assay measures compound and/or proteolysis-targeting chimera (PROTAC) binding in live cells, where cellular factors such as the plasma membrane can impact binding potency. PROTACs can have high molecular weights and suffer from poor cell permeability. To determine the permeability of novel PROTACs in a simple work flow, NanoBRET™ TE Assays can also be performed in permeabilized cells to assess compound binding to the E3 ligase in the absence of the cell membrane, which can be particularly important for understanding PROTAC permeability.