Status:
ReviewerMember since: 2014
Organization: Aurelia Bioscience
Great reagents, simple to use, excellent communication from the manufacturer
Application Area: Measurement of kinase compound binding and residency time in living cells at physiological ATP conc.
"NanoBRET TE is the first real-time biophysical method to enable quantitative assessment of kinase-inhibitor binding and residence time under physiological ‘live-cell’ conditions. The quantitative BRET-based capability is achieved via energy transfer from cell-permeable fluorescent tracers reversibly engaged to NanoLuc luciferase-tagged kinase proteins. Over 250 full length kinase have been tagged with Nanoluc by Promega creating a library of kinase that can be used for competitive binding experiments in living cells. Therefore data can be obtained on cell permeability and competitive binding within a physiological environment. In addition, measurements of residency time (how long the compound is bound to the kinase) can be made. SAR is generally driven by potency with little thought given to residency time - however what is best - 1mM binding and 15 minutes residency time, or 500nM binding and 4 hour residency time? This latter statistic may also drive off target effects as many SAR programs run a small panel screen for off target kinase binding potency but do not examine residency time parameters. Both potency and residency time can be measured using this technology. Furthermore the technology can be used to identify new binding molecules to be used as warheads in PROTAC studies. NanoBRET can detect binding at type I, type II and allosteric binding sites. Therefore molecules can be designed to bind to different areas of the kinase and used as part of a PROTAC to increase degradation rate of the kinase. The NanoBRET technology has also been applied to other targets such as epigenetic (bromo domains) and others. The reagents are simple to use and come with well written protocols to allow the user to generate data in a relatively short time. This technology has far reaching implications for the detection of binding and residency time for small molecules to targets."
Status:
ReviewerMember since: 2018
Organization: University of North Carolina at Chapel Hill
Great, innovative assay!
Application Area:Drug discovery and pharmacology validation
"The NanoBRET system is a great application of FRET technology to produce an in-cell assay for drug-kinase binding, which is helpful in the development of inhibitors."
Status:
ReviewerMember since: 2018
Organization: UNC-CH
Essential and field-changing technology.
Application Area:Running kinase assays in our lab
"Easy to use, high quality and reproducible results. Highly recommended to others!"
Status:
ReviewerMember since: 2018
Organization: UNC Eshelman School of Pharmacy
The NanoBRET cellular kinase engagement assay is poised to set a new standard!
Application Area:Kinase probe discovery in open access science
"I am thrilled with the NanoBRET cellular target engagement assay for kinases. It is easy to use, highly relevant and reproducible, and wonderfully supported by Promega."
Status:
ReviewerMember since: 2018
Organization: UNC Chapel Hill
Another highly useful product from the group at Promega !
Application Area:To better understand target engagement of kinase inhibitors
"Innovative and effective use of bioluminescent technology to help us understand how compounds interact with kinases in cells. Methodology well worked out, straightforward to use and interpret data - great tool to help study target engagement !"
Status:
ReviewerMember since: 2018
Organization: UNC
Great product. Innovative company
Application Area:Kinase assays
"Easy to use. High quality data. Reproducible. Good customer support"
Status:
ReviewerMember since: 2018
Organization: Theravance Biopharma
Great assay system
Application Area:This assay helped a lot for high throughput screening for kinase protein inhibitor.
"Can easily generate IC50 - HTP compound screening - Easy to use"
Status:
ReviewerMember since: 2018
Organization: The Institute of Cancer Research
A very exciting technique, I think it will be used more and more in the future
Application Area:TE Assay
"It gave good robust results and it was easy to use. I would recommend it."
Status:
ReviewerMember since: 2018
Organization: Roche
Fantastic tool. Let your creativity loose to discover ways it can be utilized.
Application Area:Target engagement
"NanoBRET has addressed a long-standing issue in small molecule drug discovery: what target occupancy is achieved in the complex intracellular environment."
Status:
ReviewerMember since: 2018
Organization: Reagent Genie Ireland LTD
Excellent
Application Area:Kinase screen
"Excellent product that was very easy to use. Yes, definitely recommend."
Status:
ReviewerMember since: 2018
Organization: Howard University
Best research product of the last year.
Application Area:cell signaling
"very easy to use and very effective in determining cell signaling."
Status:
ReviewerMember since: 2018
Organization: Goethe University / Structural Genomics Consortium Frankfurt
This will be the next profiling technology of future drug discovery programs!
Application Area:Focus: Protein kinase inhibitor profiling in living cells at cellular [ATP] using full-length constr
"Very easy to use due to detailed protocols from the manufacturer. Gained insights in kinase inhibitor affinities in an in vivo context at cellular [ATP] using full-length constructs. Assay z' is usually very high because of little variability between replicates. Advantage of readout over FRET: no photobleaching during data collection because of the bioluminescence readout. The instrument enables in vivo kinetic measurements in intact human cells. Would recommend."
Status:
ReviewerMember since: 2018
Organization: galapagos
Great and innovative technology
Application Area:drug discovery
"This technology enables us to test binding properties of small molecules to numerous targets in a less or more generic way. The technology was easily implemented in our screening cascade without excessive assay development. I would strongly recommend to researchers in the area of drug discovery to consider this technology."
Status:
ReviewerMember since: 2018
Organization: C4 Therapeutics
Great assay for measuring compound binding in cells.
Application Area:Measruing the cpd binding of the target protein in cells.
"The assay development process was very straight forward and Promega gave great technical support during the process. The assay itself was easy to use with high-quality results and we have implemented this assay to our current profiling workflow for 6 months now."
Status:
ReviewerMember since: 2012
Organization: Lead Discovery Center GmbH
Great product, gives new capabilities for hard to express kinase targets.
Application Area:The NanoBRET™ TE intracellular kinase assay is used for the characterization of compounds.
"The NanoBRET™ TE intracellular kinase assay protocols are easy to establish and give excellent data reproducibility. I can fully recommend the assay to others."
Status:
ReviewerMember since: 2011
Organization: AstraZeneca
Enabling technology for our applications.
Application Area:Target engagement and cell-kinetics
"The technology has been enabling for investigation of target engagement and preliminary studies of in cell-kinetics. High quality results are readily obtainable for the targets we have studied to date and the support from the R&D team at Promega has been outstanding."
Status:
ReviewerMember since: 2015
Organization: The University of North Carolina at Chapel Hill
The NanoBret has proven to be a key technology for our drug discovery efforts!
Application Area:I am using this product to evaluate target engagement of protein kinases involved in ER stress.
"Our studies are demonstrating how helpful the NanoBret technology can be for our research. It is easy to transfect cells with this reagent and the resulting data were of high quality."
Status:
ReviewerMember since: 2017
Organization: Leveragen
Great instrument.
Application Area:Lead Discovery
"It is very easy to use, time saving, and always generates reliable data."
Status:
ReviewerMember since: 2017
Organization: Galapagos NV
Great technology, easy to use!
Application Area:Hit profiling.
"NanoBRET kinase assays have been proven to be useful in most cases to add in a hit profiling cascade between biochemical assay and a cellular activity assay. The technology is user friendly."
Status:
ReviewerMember since: 2017
Organization: Bristol-Myers Squibb
Fantastic product - completely transformed our workflow!
Application Area:Single Cell ICP-MS analysis for nanotoxicity and during drug development.
"NanoBRET is a super easy-to-use kit, and we get very clean high quality data every time."
Measure Kinase-Ligand Affinity in Live Cells
The NanoBRET™ Target Engagement (TE) Intracellular Kinase Assay measures compound binding at select target kinases in intact cells. The assay is based on the NanoBRET™ System, an energy transfer technique designed to measure molecular proximity in living cells. The NanoBRET™ TE Assay measures the apparent affinity of test compounds by competitive displacement of the NanoBRET™ tracer, reversibly bound to a NanoLuc® luciferase-kinase fusions in cells.
How it Works
Using the NanoBRET™ TE Kinase Assay, a fixed concentration of tracer (K-4 or K-5) is added to cells expressing the desired NanoLuc® kinase fusion to generate a BRET signal. Introduction of competing compounds results in a dose-dependent decrease in NanoBRET™ signal, which allows quantitation of the intracellular affinity of the target protein for the test compound.
Manufacturer Promega Corp. | Available Worldwide
4.7 / 5.0 | 22 reviews