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NanoBRET™ Signaling Protein Assays

Sensitive Live Cell Assays for Screening Cell Signaling Protein Interactions

  • Order NanoBRET™ KRas/BRaf Interaction Assay by selecting N1880
  • For additional signaling assays go to "View Prebuilt Assays" link below

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Catalog number selected: N1880

$ 5,000.00
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NanoBRET™ Signaling Protein Assays
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Study Proteins in Their Natural Cellular Context

NanoBRET™ Signaling Protein Assays are sensitive, reproducible live cell assays designed for monitoring or screening the interaction of proteins involved in cell signaling events. Interactions between proteins are key events in normal cellular signal transduction pathways, and modulation of these interactions has been implicated in disease formation, making them important candidates for drug targeting. The NanoBRET™ KRas/BRaf Interaction Assay measures the specific interaction between mutant KRas (G12C) and BRaf human proteins in their natural cellular context. In epidermal growth factor receptor (EGFR) pathway-associated oncogenesis, mutations in KRas result in constitutive binding to BRaf even in the absence of growth factor, resulting in cell proliferation and suppressed apoptosis.

Better Separation Between Donor and Acceptor Signals

Energy Transfer from NanoLuc to HaloTag NanoBRET Ligand
Depiction of energy transfer from a NanoLuc®-Protein A fusion (energy donor) to a fluorescently labeled HaloTag®-Protein B fusion (energy acceptor) upon interaction of Protein A and Protein B. 
Donor and Acceptor Signal Wavelengths for NanoBRET
Spectral separation of the NanoLuc® emission (460nm) and the fluorescent HaloTag® NanoBRET™ 618 Ligand emission (618nm), and calculation of the NanoBRET™ ratio.

Notes

  • Sufficient NanoBRET™ Nano-Glo® Substrate and HaloTag® NanoBRET™ 618  Ligand are provided to perform 400 assays in 96-well plates or 1,000 assays in 384-well plates.
  • To perform NanoBRET™ PPI Assays an instrument, such as the GloMax® Discover, capable of sequentially measuring dual-filtered luminescence and equipped with appropriate filters is required.
  • The ideal filter setup will include a band pass (BP) filter centered around 460nm to measure the donor signal (e.g., Emission 450nm/BP 80nm) and a long pass (LP) filter starting at around 600–610nm to measure the acceptor signal (e.g., Emission 610nm/LP). Filters outside of these ranges will miss critical measurements and compromise data quality.
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.

Obtain High Assay Specificity

KRas-HaloTag and BRaf-NanoLuc NanoBRET DSA
Donor saturation assay performed by transfecting cells with a constant amount of BRaf-NanoLuc® donor DNA and increasing amounts of KRas(G12C)-HaloTag® acceptor DNA to represent increasing acceptor-to-donor (A:D) ratios. The negative control is membrane-localized NanoLuc® protein as a source of a mock donor in place of the BRaf-NanoLuc® fusion. The assay generates a hyperbolic curve, which indicates detection of a specific BRET interaction.
Signaling Protein Assay Vectors
12743MA-W
KRas(G12C)-HaloTag® Fusion Vector sequence & reference points.
12744MA-W
BRaf-NanoLuc® Fusion Vector sequence & reference points.
NanoBRET™ Control Vectors
12729MA-W
NanoBRET™ Positive Control Vector sequence & reference points.
12730MA-W
p53-HaloTag® Fusion Vector sequence & reference points.
12731MA-W
NanoLuc®-MDM2 Fusion Vector sequence & reference points.

Specifications

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

KRas(G12C)-HaloTag® Fusion Vector

N177A 1 × 20μg

BRaf-NanoLuc® Fusion Vector

N178A 1 × 20μg

SDS

Choose language:

Download SDSPDF (181 KB) – English (United States)

Certificate of Analysis

See all certificates for N1880

Use Restrictions

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

Storage Conditions

BB

Store at –20°C.

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:
(a) use NanoBRET™-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(b) 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. Nos. 7,425,436, 7,935,803, 8,466,269, 8,742,086, 8,420,367 and 8,748,148 and other patents and patents pending.

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.

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.

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