NanoBRET® Target Engagement BET BRD Assays
Directly Measure BET BRD-Test Compound Affinity and Residence Time
- Compound and tracer compete directly for the binding site
- Assays rely on full-length proteins that are similar to the native forms
- Express target proteins at low levels
Catalog Number:
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Size
Catalog Number: N2131
Catalog Number: N2180
Catalog Number: N2130
Catalog Number: N2140
NanoBRET® Technology for Target Engagement
The NanoBRET® Target Engagement (TE) Assay measures compound binding at select target proteins in intact cells, in real time. The NanoBRET® TE Assay uses four key components: An expressed cellular target protein that is fused to the bright NanoLuc® luciferase; a cell-permeable fluorescent tracer that specifically binds to the target protein; a substrate for NanoLuc® luciferase; and a cell-impermeable inhibitor for NanoLuc® luciferase.
The assay uses bioluminescence resonance energy transfer (BRET), achieved by transferring the luminescent energy from NanoLuc® luciferase to the fluorescent tracer that is bound to the target protein-NanoLuc® fusion. This energy transfer makes it possible to directly measure compound binding affinity as well as compound-target residence time.
Compounds that are applied to the cells and specifically engage the intracellular target protein-NanoLuc® fusion will result in a decrease in BRET. To ensure accurate assessment of intracellular target engagement, a NanoLuc® inhibitor is used to mitigate any extracellular NanoLuc® signal that may arise from cells compromised during handling, while not adversely affecting NanoLuc® luciferase expressed within healthy living cells
Analyze Binding Affinity and Permeability
The NanoBRET® TE Assay analyzes the apparent binding affinity and permeability of test compounds by competitive displacement of a NanoBRET® Tracer reversibly bound to a NanoLuc® fusion protein in cells.
The optimal NanoBRET® Tracer affinity is determined for each target protein.
The optimal NanoBRET™®Tracer affinity is determined for the bromodomain (BRD)-NanoLuc® luciferase fusion protein.
Directly Measure Residence Time
The NanoBRET® TE Assay directly measures the durability of compound-target engagement after binding in real time and provides information on full-length protein targets.
Evaluate Key Drug Target Class
The NanoBRET® Target Engagement Assay can be used to evaluate binding of compounds to BRDs, a key drug target class.
DNA Bundle Components
Protocols
Specifications
Catalog Number:
What's in the box
| Item | Part # | Size | Available Separately |
|---|---|---|---|
Transfection Carrier DNA |
E488B | 1 × 100μg | View Product |
NanoBRET® Nano-Glo® Substrate |
N157C | 1 × 330μl | View Product |
NanoLuc®-BRD4 FL Fusion Vector |
N169A | 1 × 20μg | |
Tracer Dilution Buffer |
N219A | 1 × 5ml | View Product |
NanoBRET® Intracellular TE BET BRD Tracer |
N234B | 1 × 1.1ml | |
Extracellular NanoLuc® Inhibitor |
N235B | 1 × 110μl | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no commercial use is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the product. No other use or transfer of this product is authorized without the prior express written consent of Promega.
For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(i) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(ii) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene. No other use of this product is authorized without the prior express written consent of Promega.
In addition, researcher must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product; or
(1b) contact Promega to obtain a license for use of the product with LARs not manufactured by Promega.
For uses of this product (Nano-Glo® ORF Vector) for energy transfer (such as bioluminescence resonance energy transfer), researcher must:
(2a) use NanoBRET®-branded luminescent assay reagents (LARs; e.g., NanoBRET® Nano-Glo® Substrate) or Intracellular TE Nano-Glo® Substrate/Inhibitor for all determinations of luminescent activity; and
(2b) use NanoBRET®-branded energy acceptors (e.g., NanoBRET® tracers, NanoBRET® dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity; or
(2c) contact Promega to obtain a license for use of the product for energy transfer assays using energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. Nos. 9,056,885, 10,139,400 and 10,215,751; European Pat. No. 2782916; and Japanese Pat. Nos. 6522339, 6574821 and 7227741.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
U.S. Pat. No. 10,024,862; Japanese Pat. No. 7676487; and other patents and patents pending.
What's in the box
| Item | Part # | Size |
|---|---|---|
NanoBRET® TE Intracellular BET BRD Assay |
N2131 | 1 × 1,000 assays |
NanoBRET® TE BET BRD DNA Bundle |
N2150 | 1 × 1 each |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no commercial use is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the product. No other use or transfer of this product is authorized without the prior express written consent of Promega.
For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(i) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(ii) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene. No other use of this product is authorized without the prior express written consent of Promega.
In addition, researcher must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product; or
(1b) contact Promega to obtain a license for use of the product with LARs not manufactured by Promega.
For uses of this product (Nano-Glo® ORF Vector) for energy transfer (such as bioluminescence resonance energy transfer), researcher must:
(2a) use NanoBRET®-branded luminescent assay reagents (LARs; e.g., NanoBRET® Nano-Glo® Substrate) or Intracellular TE Nano-Glo® Substrate/Inhibitor for all determinations of luminescent activity; and
(2b) use NanoBRET®-branded energy acceptors (e.g., NanoBRET® tracers, NanoBRET® dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity; or
(2c) contact Promega to obtain a license for use of the product for energy transfer assays using energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. Nos. 9,056,885, 10,139,400 and 10,215,751; European Pat. No. 2782916; and Japanese Pat. Nos. 6522339, 6574821 and 7227741.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
U.S. Pat. No. 10,024,862; Japanese Pat. No. 7676487; and other patents and patents pending.
What's in the box
| Item | Part # | Size | Concentration | Available Separately |
|---|---|---|---|---|
Transfection Carrier DNA |
E488A | 1 × 20μg | 1μg/μl | View Product |
NanoBRET® Nano-Glo® Substrate |
N157A | 1 × 50μl | View Product | |
NanoLuc®-BRD4 FL Fusion Vector |
N169A | 1 × 20μg | ||
Tracer Dilution Buffer |
N219A | 1 × 5ml | View Product | |
NanoBRET® Intracellular TE BET BRD Tracer |
N234A | 1 × 110μl | ||
Extracellular NanoLuc® Inhibitor |
N235A | 1 × 17μl | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no commercial use is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the product. No other use or transfer of this product is authorized without the prior express written consent of Promega.
For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(i) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(ii) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene. No other use of this product is authorized without the prior express written consent of Promega.
In addition, researcher must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product; or
(1b) contact Promega to obtain a license for use of the product with LARs not manufactured by Promega.
For uses of this product (Nano-Glo® ORF Vector) for energy transfer (such as bioluminescence resonance energy transfer), researcher must:
(2a) use NanoBRET®-branded luminescent assay reagents (LARs; e.g., NanoBRET® Nano-Glo® Substrate) or Intracellular TE Nano-Glo® Substrate/Inhibitor for all determinations of luminescent activity; and
(2b) use NanoBRET®-branded energy acceptors (e.g., NanoBRET® tracers, NanoBRET® dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity; or
(2c) contact Promega to obtain a license for use of the product for energy transfer assays using energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. Nos. 9,056,885, 10,139,400 and 10,215,751; European Pat. No. 2782916; and Japanese Pat. Nos. 6522339, 6574821 and 7227741.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
U.S. Pat. No. 10,024,862; Japanese Pat. No. 7676487; and other patents and patents pending.
What's in the box
| Item | Part # | Size | Available Separately |
|---|---|---|---|
NanoBRET® Nano-Glo® Substrate |
N157D | 1 × 3.3ml | View Product |
Tracer Dilution Buffer |
N219B | 1 × 50ml | View Product |
NanoBRET® Intracellular TE BET BRD Tracer |
N234B | 10 × 1.1ml | |
Extracellular NanoLuc® Inhibitor |
N235C | 1 × 1.1ml | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
See Protocol for storage recommendations.
BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If researcher is not willing to accept the terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researcher may use this product for research use only; no commercial use is allowed. Commercial use means any and all uses of this product by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; (2) use in provision of services, information or data; and (3) resale of the product, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the product. No other use or transfer of this product is authorized without the prior express written consent of Promega.
For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(i) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(ii) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.
With respect to any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, please contact Promega for supply and licensing information. PROMEGA MAKES NO REPRESENTATIONS OR WARRANTIES OF ANY KIND, EITHER EXPRESSED OR IMPLIED, INCLUDING FOR MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE, WITH REGARD TO THE PRODUCT. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. Nos. 9,056,885, 10,139,400 and 10,215,751; European Pat. No. 2782916; and Japanese Pat. Nos. 6522339, 6574821 and 7227741.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
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