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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

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Catalog number selected: N1830

$ 5,000.00
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This product is discontinued
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NanoBRET™ Bromodomain/Histone Interaction Assays
BRD4/Histone H3.3
$ 5,000.00
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32612632-L1-NanoBRET-PPI-Overview-Image

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. 

Energy Transfer from NanoLuc to HaloTag NanoBRET Ligand
Donor and Acceptor Signal Wavelengths for NanoBRET

Example NanoBRET™ BRD/Histone Protein Interaction Assay Data

NanoBRET™ BRD4 Assay
NanoBRET™ BRD9 Assay
NanoBRET™ BRPF1 Assay

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.

BRD4 Interaction with Histone H3.3 in HEK293 Cells
BRD4 Interaction with Histone H3.3 in HCT116 Cells

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.

NanoLuc-BRD9 FL and HaloTag-Histone H3.3 DSA
Donor saturation assay performed by transfecting cells with a constant amount of BRD9-NanoLuc® Fusion donor DNA.
NanoLuc-BRD9 BD and HaloTag-Histone H3.3 DSA
Donor saturation assay performed by transfecting cells with a constant amount of BRD9-NanoLuc® Fusion donor DNA

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.

NanoLuc-BRPF1 FL and HaloTag-Histone H4 Donor Saturation
Donor saturation assay performed by transfecting cells with a constant amount of BRPF1-NanoLuc® Fusion donor DNA.
NanoLuc-BRPF1 BD and HaloTag-Histone H4 Donor Saturation
Donor saturation assay performed by transfecting cells with a constant amount of BRPF1-NanoLuc® Fusion donor DNA
36488360-helpsection-MG_3480

Choose Starter Systems, Prebuilt or Custom NanoBRET™ Assays

Configure NanoBRET™ PPI Assays in a format that suits your needs. Build your own assays using the starter systems, choose from prebuilt assays in our catalog or consult with us to design a customized assay specific for your research.

Engineered Vectors Simplify Your Work

NanoLuc®-BRD4 Vectors
12900MA-W
NanoLuc®-BRD4 BD1 Fusion Vector sequence text file.
12737MA-W
NanoLuc®-BRD4 FL Fusion Vector sequence text file.
NanoLuc®-BRD9 Vectors
12741MA-W
NanoLuc®-BRD9 BD Fusion Vector sequence text file.
12740MA-W
NanoLuc®-BRD9 FL Fusion Vector sequence text file.
NanoLuc®-BRPF1 Vectors
12742MA-W
NanoLuc®-BRPF1 BD Fusion Vector sequence text file.
12899MA-W
NanoLuc®-BRPF1 FL Fusion Vector sequence text file.
NanoBRET™ Histone Vectors
12691MA-W
Histone H3.3-HaloTag® Fusion Vector sequence text file.
12692MA-W
Histone H4-HaloTag® Fusion Vector sequence text file.
NanoBRET™ Control Vectors
12729MA-W
NanoBRET™ Positive Control Vector sequence text file.
12730MA-W
p53-HaloTag® Fusion Vector sequence text file.
12731MA-W
NanoLuc®-MDM2 Fusion Vector sequence text file.

Specifications

You are viewing: N1830 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

HaloTag® NanoBRET™ 618 Ligand

G980A 2 × 20μl View Product

NanoBRET™ Nano-Glo® Substrate

N157A 2 × 50μl

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

Choose language:

Download SDSPDF (206 KB) – English (United States)

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

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. 7,867,726 and other patents and patents pending.

U.S. Pat. No. 8,809,529, European Pat. No. 2635582 and other patents and patents pending.

U.S. Pat. Nos. 7,425,436, 7,935,803, 8,466,269, 8,742,086, 8,420,367, 8,748,148, 9,416,353, 9,593,316 and other patents and patents pending.

Patent Pending.

U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730 and 9,951,373 and other patents and patents pending.

U.S. Pat. Nos. 10,067,149 and 10,024,862 and other patents and patents pending.

U.S. Pat. Nos. 9,056,885 and 10,139,400, European Pat. No. 2782916 and other patents pending.

Licensed under EP1295121 and EP1088233.

Specifications

You are viewing: N1890 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

HaloTag® NanoBRET™ 618 Ligand

G980A 2 × 20μl View Product

NanoBRET™ Nano-Glo® Substrate

N157A 2 × 50μl

p53-HaloTag® Fusion Vector

N159A 1 × 20μg

NanoLuc®-MDM2 Fusion Vector

N160A 1 × 20μg

Histone H4-HaloTag® Fusion Vector

N168A 1 × 20μg

NanoLuc®-BRD4 FL Fusion Vector

N169A 1 × 20μg

NanoLuc®-BRD4 BD1 Fusion Vector

N170A 1 × 20μg

SDS

Choose language:

Download SDSPDF (206 KB) – English (United States)

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: N1840 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

HaloTag® NanoBRET™ 618 Ligand

G980A 2 × 20μl View Product

NanoBRET™ Nano-Glo® Substrate

N157A 2 × 50μl

p53-HaloTag® Fusion Vector

N159A 1 × 20μg

NanoLuc®-MDM2 Fusion Vector

N160A 1 × 20μg

Histone H3.3-HaloTag® Fusion Vector

N167A 1 × 20μg

NanoLuc®-BRD9 FL Fusion Vector

N173A 1 × 20μg

NanoLuc®-BRD9 BD Fusion Vector

N174A 1 × 20μg

SDS

Choose language:

Download SDSPDF (204 KB) – English (United States)

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: N1900 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

HaloTag® NanoBRET™ 618 Ligand

G980A 2 × 20μl View Product

NanoBRET™ Nano-Glo® Substrate

N157A 2 × 50μl

p53-HaloTag® Fusion Vector

N159A 1 × 20μg

NanoLuc®-MDM2 Fusion Vector

N160A 1 × 20μg

Histone H4-HaloTag® Fusion Vector

N168A 1 × 20μg

NanoLuc®-BRD9 FL Fusion Vector

N173A 1 × 20μg

NanoLuc®-BRD9 BD Fusion Vector

N174A 1 × 20μg

SDS

Choose language:

Download SDSPDF (204 KB) – English (United States)

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: N1860 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

HaloTag® NanoBRET™ 618 Ligand

G980A 2 × 20μl View Product

NanoBRET™ Nano-Glo® Substrate

N157A 2 × 50μl

p53-HaloTag® Fusion Vector

N159A 1 × 20μg

NanoLuc®-MDM2 Fusion Vector

N160A 1 × 20μg

Histone H3.3-HaloTag® Fusion Vector

N167A 1 × 20μg

NanoLuc®-BRPF1 FL Fusion Vector

N175A 1 × 20μg

NanoLuc®-BRPF1 BD Fusion Vector

N176A 1 × 20μg

SDS

Choose language:

Download SDSPDF (205 KB) – English (United States)

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: N1910 Change Configuration

What's in the box?

Item Part # SizeAvailable Separately

HaloTag® NanoBRET™ 618 Ligand

G980A 2 × 20μl View Product

NanoBRET™ Nano-Glo® Substrate

N157A 2 × 50μl

p53-HaloTag® Fusion Vector

N159A 1 × 20μg

NanoLuc®-MDM2 Fusion Vector

N160A 1 × 20μg

Histone H4-HaloTag® Fusion Vector

N168A 1 × 20μg

NanoLuc®-BRPF1 FL Fusion Vector

N175A 1 × 20μg

NanoLuc®-BRPF1 BD Fusion Vector

N176A 1 × 20μg

SDS

Choose language:

Download SDSPDF (205 KB) – English (United States)

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

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