Nano-Glo® HiBiT Dual-Luciferase® Reporter System
Multiplexed Detection of HiBiT-Tagged Proteins and Firefly Luciferase in the Same Sample
- Rapidly identify specific and non-specific effects of compounds
- Compatible with batch processing and automation
Catalog Number:
Size
Catalog Number: NE2010
Catalog Number: NE2020
Sequential Measurement of Firefly Luciferase and HiBiT-Tagged Proteins from the Same Sample
The Nano-Glo® HiBiT Dual-Luciferase® Reporter System (HiBiT NanoDLR™) allows sequential measurement of firefly luciferase (Fluc) and HiBiT-tagged proteins from the same sample in an “add-read-add-read” assay format. Use Fluc as a constitutively expressed control to indicate nonspecific compound effects on cell viability or protein expression, rather than specific changes in the levels of the HiBiT-tagged protein of interest, aiding in the interpretation of protein modulation screens and studies.
Simplify Your Workflow
Explore premade solutions to simplify your workflow—whether you need ready-to-use firefly expression vectors, HiBiT-tagged protein vectors or prebuilt knock-in CRISPR cell lines designed for reliable expression and detection.
HiBiT NanoDLR™ Workflow
Firefly luciferase (Fluc) is first detected after addition of the ONE-Glo™ EX reagent supplemented with LgBiT (1). The NanoDLR™ Stop & Glo® Reagent is added to the cells to both quench the firefly signal and detect the now-complemented NanoBiT® (HiBiT + LgBiT) signal (2).
Use the Fluc Signal to Distinguish Specific and Nonspecific Compound Effects on Protein Levels
Selective HiBiT-BRD4 degradation with two PROTACs compared to general cellular toxicity. HEK293 cells were transiently transfected with a construct expressing the HiBiT-BRD4-IRES-luc2 cassette from the weak TK promoter. Cells were treated with titrating concentrations of MZ1 (Panel A), dBET1 (Panel B) or staurosporine (Panel C) before the Fluc (blue) and HiBiT (red) signals were measured using the lytic HiBiT NanoDLR™ Assay. MZ1 and dBET1 selectively link BRD4 to two different E3 ligase complexes for targeted degradation, and staurosporine causes general toxicity, resulting in loss of both HiBiT-BRD4 and firefly luciferase (Fluc).
Quantifiable Over Seven Orders of Magnitude
The broad linear dynamic range accurately quantifies tagged proteins regardless of expression level to measure changes in protein abundance. The HiBiT NanoDLR™ Assay can quantify even low-abundance proteins at endogenous levels of expression.
Protocols
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
ONE-Glo™ EX Luciferase Assay Buffer |
E632A | 1 × 10ml |
ONE-Glo™ EX Luciferase Assay Substrate |
E633A | 1 × 1 vial |
NanoDLR™ Stop & Glo® Buffer |
N142A | 1 × 10ml |
NanoDLR™ Stop & Glo® Substrate |
N143A | 1 × 100μl |
LgBiT Protein |
N401A | 1 × 0.1ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
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. 9,487,814, European Pat. No. 2959010 and other patents and patents pending.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. No. 9,797,889 and other patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
ONE-Glo™ EX Luciferase Assay Buffer |
E632B | 1 × 100ml |
ONE-Glo™ EX Luciferase Assay Substrate |
E633B | 1 × 1 vial |
NanoDLR™ Stop & Glo® Buffer |
N142B | 1 × 100ml |
NanoDLR™ Stop & Glo® Substrate |
N143B | 1 × 1ml |
LgBiT Protein |
N401B | 1 × 1ml |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Storage Conditions
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. 9,487,814, European Pat. No. 2959010 and other patents and patents pending.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. No. 9,797,889 and other patents pending.
Resources
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