Promega, NP5000, NanoBRET® Target Engagement CDK Selectivity Systems

Profile Compound Selectivity for Cyclin-Dependent Kinases (CDKs) in Live Cells
Determine compound or inhibitor affinity and occupancy against up to 16 CDKs
Each NanoBRET® TE CDK Selectivity System uses a simple, addition-only microplate workflow and provides a CDK Vector Panel (A or B) and a NanoBRET® TE Kinase Assay
Two transfection-ready NanoBRET® TE CDK vector panels (A and B) with 8 individual pairings of CDK-NanoLuc® fusion vector with cyclin/carrier DNA
Specifications：
CDK1/CCNB1 Vector Mix NP501A 1 × 120μl
CDK2/CCNE1 Vector Mix NP502A 1 × 120μl
CDK3/CCNE1 Vector Mix NP503A 1 × 120μl
CDK4/CCND3 Vector Mix NP504A 1 × 120μl
CDK5/CDK5R1 Vector Mix NP505A 1 × 120μl
CDK6/CCDN1 Vector Mix NP506A 1 × 120μl
CDK7/Carrier Vector Mix NP507A 1 × 120μl
CDK9/CCNK Vector Mix NP508A 1 × 120μl
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
NanoBRET® Tracer K-10 N264B 1 × 300μl
LIMK1-NanoLuc® Fusion Vector NV339A 1 × 20μg
CDK1/CCNB1 Vector Mix NP501A 1 × 120μl
CDK2/CCNE1 Vector Mix NP502A 1 × 120μl
CDK3/CCNE1 Vector Mix NP503A 1 × 120μl
CDK4/CCND3 Vector Mix NP504A 1 × 120μl
CDK5/CDK5R1 Vector Mix NP505A 1 × 120μl
CDK6/CCDN1 Vector Mix NP506A 1 × 120μl
CDK7/Carrier Vector Mix NP507A 1 × 120μl
CDK9/CCNK Vector Mix NP508A 1 × 120μl
CDK10/CCNL2 Vector Mix NP511A 1 × 120μl
CDK14/CCNY Vector Mix NP512A 1 × 120μl
CDK15/CCNY Vector Mix NP513A 1 × 120μl
CDK16/CCNY Vector Mix NP514A 1 × 120μl
CDK17/CCNY Vector Mix NP515A 1 × 120μl
CDK18/CCNY Vector Mix NP516A 1 × 120μl
CDKL2/Carrier Vector Mix NP517A 1 × 120μl
CDK20/CCNH Vector Mix NP518A 1 × 120μl
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
NanoBRET® Tracer K-10 N264B 1 × 300μl
LIMK1-NanoLuc® Fusion Vector NV339A 1 × 20μg
CDK10/CCNL2 Vector Mix NP511A 1 × 120μl
CDK14/CCNY Vector Mix NP512A 1 × 120μl
CDK15/CCNY Vector Mix NP513A 1 × 120μl
CDK16/CCNY Vector Mix NP514A 1 × 120μl
CDK17/CCNY Vector Mix NP515A 1 × 120μl
CDK18/CCNY Vector Mix NP516A 1 × 120μl
CDKL2/Carrier Vector Mix NP517A 1 × 120μl
CDK20/CCNH Vector Mix NP518A 1 × 120μl
The NanoBRET® Target Engagement (TE) CDK Selectivity Systems quantitatively measure intracellular compound affinity and occupancy against two panels of 8 CDKs each, allowing you to measure up to 16 CDKs in a single experiment. Each vector panel contains 8 individual tubes of CDK-NanoLuc® fusion vectors that are premixed with either a cyclin expression vector or carrier DNA for optimal transfections.
The CDK Selectivity Systems rely on the
NanoBRET® TE technology
, a bioluminescence resonance energy transfer (BRET) method that uses the small, extremely bright NanoLuc® luciferase as the energy donor and a cell-permeable NanoBRET® TE Tracer as the energy acceptor. The CDK Selectivity Systems use an addition-only, multiwell plate format with a single NanoBRET® TE Kinase Assay.
In this study, the NanoBRET® TE method was used to profile a collection of 46 clinically advanced CDK inhibitors and tool molecules in live cells. The results confirmed the binding of advanced clinical compounds and revealed new drug-CDK interactions. These new findings offer the potential to repurpose drugs for lesser-studied CDK family members. This paper highlights the importance of determining drug selectivity across the CDK family in live cells. The NanoBRET® TE CDK Selectivity Systems expanded on this work, simplifying the workflow to use a single NanoBRET® TE Assay (K-10) and ready-to-use CDK Vector Panels.
Wells, C.I.
et al.
(2020) Quantifying CDK inhibitor selectivity in live cells.
Nat. Comm.
11
, 2743.