Promega, N2751, NanoBRET® TE E3 Ligase Vectors

NanoLuc® E3 Ligase Fusion Vectors and DDB1 Expression Vectors for NanoBRET® TE E3 Ligase Assays
Transfection-ready vectors to express NanoLuc® luciferase fused to E3 ligase CRBN or VHL proteins
Transfection-ready vector to express DDB1 protein
Use these vectors with NanoBRET® TE Intracellular E3 Ligase Assays, CRBN or VHL, to measure E3 ligase target engagement
Specifications：
NanoLuc®-CRBN Fusion Vector N274A 1 × 20μg
VHL-NanoLuc® Fusion Vector N275A 1 × 20μg
DDB1 Expression Vector N276A 3 × 20μg
Protein degradation via small molecules such as PROTACs or molecular glues is governed by multiple steps. The initial steps involve entry of the small molecule degrader compound into the cell, followed by binding to the E3 ubiquitin ligase and protein target. The
NanoBRET® Target Engagement (TE) Intracellular E3 Ligase Assays
allow live-cell measurement of compound binding or affinity for an E3 ubiquitin ligase protein, Cereblon (CRBN) or von Hippel-Lindau disease tumor suppressor (VHL). In addition, the assays can be used to quantify intracellular availability of the compound. These assays are based on the NanoBRET® technology, an energy transfer technique designed to measure molecular proximity in living cells.
The NanoLuc® E3 ligase fusion vectors are designed to be the energy donor in the NanoBRET® TE Intracellular E3 Ligase Assay. The energy acceptor in these assays is a fluorescent cell-permeable NanoBRET® Tracer that binds to the NanoLuc® E3 ligase fusion protein. The close proximity of the tracer to NanoLuc® in the E3 ligase fusion results in a NanoBRET® signal. Addition of compounds to cells that bind the E3 ligase results in a dose-dependent decrease in NanoBRET® signal. This data allows quantitation of the intracellular affinity of degrader compounds for a target E3 ligase protein.