Promega, CS1810E23, NanoBRET® TE Intracellular E3 Ligase Assays

Measure E3 Ligase Target Engagement and Compound Intracellular Availability
Quantitate intracellular affinity of compounds for CRBN or VHL E3 ubiquitin ligases in live cells
Execute a simple workflow to calculate compound intracellular availability, a measure of permeability, to aid in compound prioritization
Scalable assays using addition-only protocols
Specifications：
NanoBRET® Nano-Glo® Substrate N157A 1 × 50μl
Tracer Dilution Buffer N219A 1 × 5ml
Extracellular NanoLuc® Inhibitor N235A 1 × 17μl
NanoLuc®-CRBN Fusion Vector N274A 1 × 20μg
DDB1 Expression Vector N276A 1 × 20μg
NanoBRET® TE Tracer CRBN N290A 1 × 30μl
NanoBRET® Nano-Glo® Substrate N157C 1 × 330μl
Tracer Dilution Buffer N219A 1 × 5ml
Extracellular NanoLuc® Inhibitor N235B 1 × 110μl
NanoLuc®-CRBN Fusion Vector N274A 1 × 20μg
DDB1 Expression Vector N276A 3 × 20μg
NanoBRET® TE Tracer CRBN N290B 1 × 300μl
NanoBRET® Nano-Glo® Substrate N157D 1 × 3.3ml
Tracer Dilution Buffer N219B 1 × 50ml
Extracellular NanoLuc® Inhibitor N235C 1 × 1.1ml
NanoBRET® TE Tracer CRBN N290B 10 × 300μl
Transfection Carrier DNA E488A 1 × 20μg
NanoBRET® Nano-Glo® Substrate N157A 1 × 50μl
Tracer Dilution Buffer N219A 1 × 5ml
Extracellular NanoLuc® Inhibitor N235A 1 × 17μl
VHL-NanoLuc® Fusion Vector N275A 1 × 20μg
NanoBRET® TE Tracer VHL N292A 1 × 30μl
Transfection Carrier DNA E488B 1 × 100μg
NanoBRET® Nano-Glo® Substrate N157C 1 × 330μl
Tracer Dilution Buffer N219A 1 × 5ml
Extracellular NanoLuc® Inhibitor N235B 1 × 110μl
VHL-NanoLuc® Fusion Vector N275A 1 × 20μg
NanoBRET® TE Tracer VHL N292B 1 × 300μl
NanoBRET® Nano-Glo® Substrate N157D 1 × 3.3ml
Tracer Dilution Buffer N219B 1 × 50ml
Extracellular NanoLuc® Inhibitor N235C 1 × 1.1ml
NanoBRET® TE Tracer VHL N292B 10 × 300μl
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® TE Intracellular E3 Ligase Assays quantitate both intracellular
target engagement and intracellular degrader compound availability.
These assays are based on the NanoBRET® technology, an energy transfer technique designed to measure molecular
proximity in living cells. S
pecifically, the NanoBRET®
Target Engagement (TE) Intracellular E3 Ligase Assays allow live cell measurement of degrader compound affinity for an E3 ubiquitin ligase protein, Cereblon (CRBN) or von Hippel-Lindau disease tumor suppressor (VHL).
I
n addition, the assays can be used to quantify intracellular availability of a compound.
As heterobifunctional degraders like PROTACs often have high molecular weight, they may suffer from poor cell
permeability or intracellular availability. By performing a simple workflow involving two unique assay modes (live-cell and permeabilized-cell),  the NanoBRET®
TE E3 Ligase Assays can be used to calculate the intracellular availability of a degrader compound.