NanoBRET® Bromodomain/Histone Interaction Assays
Protein Interaction Assays to Study BRD/Histone Interactions in Live Cells
- Study full-length bromodomain/histone-containing proteins or fragments
- Detect protein interactions in live cells, even at low expression levels
- Choose from prebuilt or custom assay options
- All but NanoBRET® BRD4/Histone H3.3 Interaction Assay are discontinued
Catalog Number:
Size
Catalog Number: N1830
Study BRD/Histone Interactions in the Natural Cellular Context
Bromodomain (BRD)-containing proteins are critical components of nuclear protein complexes involved in the recruitment of chromatin-modifying enzymes and transcriptional regulation of acetylated chromatin. The protein:protein interaction (PPI) of the BRD-containing proteins with acetylated histones is an important method of epigenetic regulation critical for cell health and development and is of great interest for drug targeting because dysfunction in BRD modulation has been implicated as a critical event in disease formation.
The NanoBRET® Bromodomain/Histone Interaction Assays enable interaction studies of bromodomain-containing proteins with full-length histones in the context of natural chromatin. In addition to the full-length bromodomain proteins, BRD fragments alone (BRD4, BRD9 and BRPF1) are also included for users that may want to understand the interaction of this isolated domain.
NanoBRET® assay technology is dependent upon energy transfer from a luminescent donor (NanoLuc® luciferase) to a fluorescent acceptor (HaloTag® NanoBRET® 618 Ligand). NanoLuc® luciferase HaloTag® protein are fused to the target proteins of interest and fusion proteins expressed at low cellular levels, enabling monitoring and screening studies of protein interactions that reflect true cellular physiology. The NanoBRET® Assay is fully reversible, enabling studies of both induction and inhibition of protein interactions.
Example NanoBRET® BRD/Histone Protein Interaction Assay Data
Example BRD4 Assay Data: Monitoring Inhibition of Protein Interactions
This example shows the dose-dependent inhibition of BRD4 protein interaction with Histone H3.3 by JQ1 as measured by the decrease in the NanoBRET®™ signal.
Example BRD9 Assay Data: High Assay Specificity
In this donor saturation assay, cells were transfected with a constant amount of BRD9 FL- or BRD9 BD-NanoLuc® Fusion donor DNA and increasing amounts of Histone H3.3-HaloTag® Fusion acceptor DNA to represent increasing acceptor-to-donor (A:D) ratios. The negative control was a set of samples containing HaloTag® protein alone. Both NanoBRET® BRD9/Histone H3.3 Interaction Assays generate hyperbolic curves, which indicate detection of a specific BRET interaction.
Example BRPF1 Assay Data: Assay Specificity
This donor saturation assay was performed by transfecting cells with a constant amount of BRPF1- or BRPF1-NanoLuc® Fusion donor DNA and increasing amounts of Histone H4-HaloTag® Fusion acceptor DNA to represent increasing amounts of acceptor-to-donor (A:D) ratios. The negative control was HaloTag® protein alone in place of the Histone H4-HaloTag® fusion. The NanoBRET® BRPF1/Histone H4 Interaction Assays generate hyperbolic curves, which indicate detection of a specific BRET interaction.
Choose Starter Systems, Prebuilt or Custom NanoBRET® Assays
Get started with NanoBRET® PPI Assays in a format that suits your needs. Design your own assays using the NanoBRET® Starter Systems, or choose from prebuilt, optimized assays for a number of popular research targets including our newest collection of RAS pathway interaction assays. You can also consult with us to design a customized assay specific for your research.
Engineered Vectors Simplify Your Work
Protocols
Complete Protocol
Specifications
Catalog Number:
Lieferumfang
| Produkt | Katalognummer | Größe | Available Separately |
|---|---|---|---|
HaloTag® NanoBRET® 618 Ligand |
G980A | 2 × 20μl | View Product |
NanoBRET® Nano-Glo® Substrate |
N157A | 2 × 50μl | View Product |
p53-HaloTag® Fusion Vector |
N159A | 1 × 20μg | |
NanoLuc®-MDM2 Fusion Vector |
N160A | 1 × 20μg | |
Histone H3.3-HaloTag® Fusion Vector |
N167A | 1 × 20μg | |
NanoLuc®-BRD4 FL Fusion Vector |
N169A | 1 × 20μg | |
NanoLuc®-BRD4 BD1 Fusion Vector |
N170A | 1 × 20μg |
SDS
Search for SDSAnalysezertifikat
Nutzungseinschränkung
For Research Use Only. Not for Use in Diagnostic Procedures.Lagerhinweise
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 or derivatives 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 or its derivatives, whether or not such product or derivatives are resold for use in research. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene or the HaloTag® gene except that researcher may create fused gene sequences provided that the coding sequence of the resulting luciferase gene or HaloTag® gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact gene sequence. No other use of this product or derivatives 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 and its derivatives, or
(1b) contact Promega to obtain a license for use of the product and its derivatives with LARs not manufactured by Promega.
For uses of Nano-Glo®-branded LARs intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researcher must:
(2a) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product and its derivatives, or
(2b) contact Promega to obtain a license for use of the product and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega.
Researcher may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license. 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 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. 8,420,367, 8,748,148, 9,593,316 and 9,873,866; European Pat. Nos. 2374875, 2492342 and 2502990; and Japanese Pat. Nos. 5680302 and 5840117.
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.
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.
Resources
No related resources found
Related Products
Ähnliche Produkte
NanoBRET® PPI Starter Systems
Live-cell protein:protein interaction assay. Includes vectors, controls and detection reagent for BRET-based PPI.
N1811, N1821
NanoBRET® Transcriptional Protein Assays
Sensitive live-cell assays for monitoring interaction of proteins involved in transcriptional regulation.
N1870
Wird oft zusammen gekauft
GloMax® Discover System
High-performance microplate reader for detecting luminescence, fluorescence and absorbance.
GM3000
FuGENE® HD Transfektionsreagenz
Verbesserte Leistung bei einer Vielzahl von Zelltypen, einschließlich schwer zu transfizierender Zelllinien.
E2311, E2312
ViaFect™ Transfection Reagent
High-efficiency, low-toxicity transfection of many cell types, including iPS cells.
E4981, E4982