Nano-Glo® HiBiT Blotting System
Fast Luminescent Detection of Proteins on Blots
- Determine protein size and quantify expression on blots
- Protocol requires only minutes, with few processing steps
- Femtogram sensitivity proportional over five orders of magnitude
Catalog Number:
Size
Rapid Detection of HiBiT-Tagged Proteins on Blots
The Nano-Glo® HiBiT Blotting System visualizes HiBiT-tagged proteins on blots at subpicogram levels. The reaction uses a detection reagent containing LgBiT Protein, which complements the HiBiT tag to form the luminescent NanoBiT® enzyme.
The blotting system requires as little as 5 minutes to detect HiBiT-tagged proteins on a nitrocellulose membrane. Standard antibody-based blotting protocols can take multiple hours to detect the protein of interest.
Simple Antibody-Free Detection Workflow
Using the Nano-Glo® HiBiT Blotting System, any HiBiT-tagged proteins can be visualized on membranes following gel separation. The blotting reagent, which contains the LgBiT Protein and furimazine substrate, is added directly to the membrane, and a luminescent signal is produced only where HiBiT is present.
This simple method requires as few as 5 minutes to perform in contrast to the multiple hours and many steps needed for standard antibody-based blotting protocols. Because luminescence is only produced only where HiBiT is present, background is minimal.
Sensitive Visualization of Blotted Proteins
HiBiT-tagged proteins can be detected at subpicogram quantities using the Nano-Glo® HiBiT Blotting System. This means the system is sensitive enough to detect proteins at endogenous expression levels.
- Verify molecular weight of HiBiT-tagged proteins
- Identify expression of splice variants
- Quickly screen transient transfections or CRISPR knock-ins
Signal Proportional Over 5 Orders of Magnitude
Confirms Expression After Transient Transfection or CRISPR Integration
HiBiT Blotting of Transiently Transfected HeLa Cells
The Nano-Glo® HiBiT Blotting System can confirm the molecular weight of HiBiT-tagged proteins.
Confirming Endogenous HiBiT Tagging of GAPDH
You can screen for effectiveness of guide RNAs and demonstrate successful editing after CRISPR/Cas9 insertion.
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
Complete Protocol
Specifications
Catalog Number:
Contenido
| Item | Part # | Presentación |
|---|---|---|
| Nano-Glo® Luciferase Assay Substrate | N113A | 1 × 200μl |
Nano-Glo® Blotting Buffer |
N242A | 1 × 10ml |
LgBiT Protein |
N401C | 1 × 0.5ml |
SDS
Search for SDSCertificado de Análisis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.Condiciones de Almacenaje
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.
U.S. Pat. No. 9,797,889 and other patents pending.
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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.
