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Nano-Glo® HiBiT Protein Quantitation System |
The HiBiT tagging system directly detects protein abundance with a simple luminescent signal. No antibodies required. Use for CRISPR knock-ins, GPCR trafficking or protein degradation studies. Choose among bioluminescent-based detection systems and HiBiT expression vectors to quantify HiBiT-tagged proteins.
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Nano-Glo® HiBiT Blotting System
Rapid Detection of HiBiT-Tagged Proteins on Blots
The Nano-Glo® HiBiT Blotting System visualizes HiBiT-tagged proteins (either tagged by cloning or CRISPR-insertion) 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.
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.
Nano-Glo® HiBiT Extracellular Detection System
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.
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
Nano-Glo® HiBiT Lytic Detection System
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
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