Nano-Glo® HiBiT Extracellular Detection System
Quantify HiBiT-Tagged Proteins Expressed on the Cell Surface
- Specific, live-cell detection of extracellular expressed or secreted proteins
- Simple add-mix-read assay format—no antibodies required
- Quantitate over 7 logs of linear dynamic range
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$ 244.00
Catalog Number: N2420
$ 1,483.00
Catalog Number: N2421
$ 11,946.00
Catalog Number: N2422
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Simple Bioluminescent Detection of Cell-Surface Protein Expression
The Nano-Glo® HiBiT Extracellular Detection System quantitates cell surface or secreted protein expression in minutes using a simple add-mix-read assay format.
Using the nonlytic Nano-Glo® HiBiT Extracellular Detection Reagent, any proteins that are tagged with the 11 amino acid HiBiT peptide and expressed outside of the cell can be specifically quantitated. The detection reagent contains the complementary polypeptide LgBiT, which spontaneously interacts with the HiBiT tag to reconstitute the bright, luminescent NanoBiT® enzyme.
Luminescence is proportional to the amount of HiBiT-tagged protein present outside of the cell over seven orders of magnitude.
Simple Add-Mix-Read Assay Workflow
You can directly measure the amount of HiBiT-tagged protein expressed outside of the cell by adding the Nano-Glo® HiBiT Extracellular Detection Reagent, mixing with cells and measuring luminescence.
The reconstituted Nano-Glo® HiBiT Detection Reagent contains the enzyme substrate and membrane-impermeable LgBiT peptide. Only HiBiT-tagged proteins expressed on the cell surface or secreted in the culture medium will complement the extracellular LgBiT to reconstitute the luminescent NanoBiT® enzyme.
The highly quantitative Nano-Glo® HiBiT Extracellular Detection System has significantly fewer processing steps compared to standard antibody-based detection, saving you time and improving accuracy.
Develop quantitative assays for:
- Receptor internalization
- Receptor recycling
- Protein or cytokine secretion
- Surface protein trafficking
Measures Receptor Internalization in Minutes
The Nano-Glo® HiBiT Extracellular Detection System make it possible to develop simple, quantitative assays for receptor internalization that save time and eliminate the variability associated with antibody-based methods. Using the optimized detection reagent results in rapid equilibration with protein receptors that minimizes well-to-well variability and captures rapidly changing biology. For example, in the figure to the right, we measured internalization of β2-adrenergic receptor (ADRB2) when exposed to four full and partial agonists.
| Agonist | EC50 | Percentage Receptor Remaining on Surface |
|---|---|---|
| Isoproterenol | 50.9nM | 16% |
| Salbutamol | 161nM | 45% |
| Salmeterol | 1.04nM | 63% |
| Formoterol | 2.92nM | 16% |
Scale from Bench to HTS
Compatibility with multiwell plates and high-throughput processing means the Nano-Glo® HiBiT Extracellular Detection System offers more flexibility and utility. Therefore, you can develop screening assays for membrane receptor biology or protein secretion not possible with standard immunodetection methods.
Internalization of HiBiT-ADRB2 in stably transfected HEK293 cells in 384-well plates
Sensitivity to Measure Endogenous Protein Levels
The small size of the 11 aa HiBiT tag enables simple cloning-free genome editing. Using CRISPR-Cas9, the HiBiT tag can be precisely inserted at endogenous loci. Therefore, HiBiT is ideal for studying the biology of endogenously regulated membrane receptors, reducing overexpression artifacts and maintaining proper stoichiometry with interacting proteins.
Learn how to accurately measure protein surface expression with the Nano-Glo® HiBiT Extracellular and Lytic Assays in this whitepaper.
Protocols
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Extracellular Buffer |
N243A | 1 × 10ml |
Nano-Glo® HiBiT Extracellular Substrate |
N300A | 1 × 0.2ml |
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. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Extracellular Buffer |
N243B | 1 × 100ml |
Nano-Glo® HiBiT Extracellular Substrate |
N300B | 2 × 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. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Extracellular Buffer |
N243B | 10 × 100ml |
Nano-Glo® HiBiT Extracellular Substrate |
N300C | 5 × 4ml |
LgBiT Protein |
N401B | 10 × 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. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
Resources
Articles
- Illuminating Transmembrane Proteins Using HiBiT CRISPR Cell Lines
- HiBiT: A Tiny Tag Expanding Endogenous Protein Detection
- Understanding Appetite Using HiBiT Protein Tagging
- Development of a HiBiT Peptide-Based NanoBRET Ligand Binding Assay for Galanin Receptor 1 in Live Cells
- Quantifying Protein Abundance at Endogenous Levels
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Frequently Used With
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Simplify Your Workflow
Explore premade solutions to simplify your workflow—whether you need ready-to-use HiBiT-tagged protein vectors or prebuilt knock-in CRISPR cell lines designed for reliable expression and detection.
