Nano-Glo® HiBiT Lytic Detection System
Accurately Quantify Any HiBiT-Tagged Protein Expressed in Cells
- Sensitive bioluminescent protein detection
- Simple add-and-read assay—no antibodies required
- Quantitate over 7 logs of linear dynamic range
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Catalog Number: N3030
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Catalog Number: N3040
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Catalog Number: N3050
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Simple, Sensitive Bioluminescent Protein Quantitation
The Nano-Glo® HiBiT Lytic Detection System quantifies cellular protein in minutes with high sensitivity and a broad dynamic range using a simple add-mix-read assay format.
The 11 amino acid HiBiT peptide tag can be added easily to a protein of interest, and the total amount of HiBiT-tagged protein in cells measured by adding the Nano-Glo® HiBiT Lytic Detection Reagent.
The detection reagent contains the complementing polypeptide LgBiT, which spontaneously interacts with the HiBiT tag to reconstitute the bright, luminescent NanoBiT® enzyme. The luminescence intensity is directly proportional to the amount of HiBiT-tagged protein in the cell lysate over seven orders of magnitude. The glow-type luminescent signal is stable for hours.
Fast and Easy Protein Quantification–Just Add, Mix and Read
With the Nano-Glo® HiBiT Lytic Detection System, the quantity of HiBiT-tagged protein is measured directly in cell lysates with a simple add-mix-read assay. The Nano-Glo® HiBiT Detection Reagent is added to cells expressing HiBiT-tagged proteins, lysing the cells and providing the LgBiT Protein and furimazine substrate necessary for luminescence.
The high-affinity interaction between LgBiT Protein and HiBiT tag reconstitutes the luminescent NanoBiT® enzyme. This results in a highly quantitative assay with fewer processing steps compared to standard antibody-based detection methods.
Develop quantitative assays for:
- Regulated protein expression
- Targeted protein degradation
- Viral infection
- Verification of protein levels from gene expression and RNA-seq experiments
- Protein abundance following pull-downs or transient transfection
Precise Protein Quantification
The broad linear dynamic range accurately quantifies tagged proteins regardless of expression level to measure changes in protein abundance. With a limit of detection of less than 10-19 moles, the Nano-Glo® HiBiT Lytic Detection System can quantify even low abundance proteins at endogenous levels of expression.
Titrating purified HiBiT-tagged protein to demonstrate linear dynamic range
Study Proteins Under Endogenous Regulation
Using CRISPR-Cas9 gene editing, HiBiT-tagged proteins can be expressed under endogenous regulatory conditions, reducing overexpression artifacts and maintaining proper stoichiometry with other endogenous binding partners or regulatory machinery.
The small 11 aa HiBiT tag can be added to the desired protein in one of two ways:
- Traditional recombinant methods with your promoter of choice
- Insertion into endogenous loci using a simple, clone-free CRISPR-Cas9 genome editing protocol
Scalable from Bench to HTS
The simple protocol and luminescence half-life of greater than 3 hours make the Nano-Glo® HiBiT Lytic Detection System ideal for batch processing multiple plates in high-throughput applications. This results in development of HTS assays for protein regulation not possible with antibody-based methods.
Induced degradation of HiBiT-BRD4 in a CRISPR-derived pool of cells in 384-well plates
Real-World Example: Using CRISPR Gene Editing to Add HiBiT to Endogenous Genes
Learn how the tiny 11-amino-acid luminescent tag called HiBiT can quantify protein abundance even at endogenous levels. CRISPR-mediated gene editing was used add the HiBiT tag to several endogenous genes, including HIF1α, and quantified protein levels in response to various stimuli.
Protocols
Specifications
Catalog Number:
What's in the box?
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Lytic Substrate |
N246A | 1 × 0.2ml |
Nano-Glo® HiBiT Lytic Buffer |
N247A | 1 × 10ml |
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.
Patent Pending.
U.S. Pat. Nos. 9,797,889 and 9,797,890 and other patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Lytic Substrate |
N246B | 2 × 1ml |
Nano-Glo® HiBiT Lytic Buffer |
N247B | 1 × 100ml |
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.
Patent Pending.
U.S. Pat. Nos. 9,797,889 and 9,797,890 and other patents pending.
What's in the box?
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Lytic Substrate |
N246C | 5 × 4ml |
Nano-Glo® HiBiT Lytic Buffer |
N247B | 10 × 100ml |
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.
Patent Pending.
U.S. Pat. Nos. 9,797,889 and 9,797,890 and other patents pending.
Resources
Articles
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