NanoBiT® BiBiT PPI Starter System
Streamlined NanoBiT® Assays with One Plasmid and Improved Reproducibility
- Single-vector design simplifies stable cell line creation
- Real-time luminescence enables dynamic PPI detection in live cells
- High signal-to-background for sensitive, reproducible results that can be easily scaled for HTS
- Ideal for drug screening, signaling and biologics research
Catalog Number:
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Catalog Number: CS1603B393
This product is available under our Early Access program - Learn More
Catalog Number: CS1603B394
This product is available under our Early Access program - Learn More
Catalog Number: CS1603B440
This product is available under our Early Access program - Learn More
Streamlined, Sensitive Protein Interaction Assays with Single-Vector Simplicity
NanoBiT® BiBiT PPI Starter Systems use NanoBiT® (NanoLuc® Binary Technology) luciferase complementation, with LgBiT and SmBiT subunits, to monitor protein:protein interactions in live cells via luminescence. The low-affinity, reversible enzyme complementation allows dynamic observation of both PPI formation and disruption.
BiBiT-ready vectors support flexible tagging configurations and are compatible with transient or stable expression from a single vector. Notably, the system uses a CMV promoter, enabling stronger expression across a wide range of cell lines, unlike the TK promoter in earlier versions.
The system streamlines assay setup and improves consistency, making it ideal for drug discovery, signaling pathways and functional PPI studies.
BiBiT Approach to Stable Cell Line Development
Generate up to eight BiBiT-ready vectors to screen and identify the optimal tagging orientation for your proteins of interest. The BiBiT-ready vector pair providing the highest signal-to-background ratio can be ligated into a single BiBiT vector, driving stable, balanced expression of both fusion proteins via a bidirectional CMV promoter. Blasticidin resistance allows selection of stable integration events in mammalian cells.
Advantages of the BiBiT Single-Vector Approach
- Co-expression of both fusion proteins at similar levels from the same locus
- Reduced variability
- Increased signal-to-background ratio
- Enhanced assay sensitivity
Improve Assay Robustness with BiBiT-Based Stable Cell Lines
Generating stable cell lines from a BiBiT vector reduces variability and increases the reliability of NanoBiT® Assays—especially in high-throughput screening environments.
Compared to traditional transient expression methods, the BiBiT-based stable expression:
- Improves assay precision
- Enhances signal-to-background ratios
- Delivers stronger Z′ values, supporting confident compound profiling and screening
| Classic NanoBiT® Transient | NanoBiT® BiBiT Stable | |
| IC50 (nM) | 238 | 343 |
| SB | 11 | 19 |
| SN | 11 | 83 |
| Z′ | 0.68 | 0.96 |
Performance comparison of CRAF/BRAF NanoBiT® Assays using transient vs. BiBiT-based stable expression. Stable cell lines show improved Z′ and assay windows, indicating higher assay quality and reduced variability.
Quantify Antibody Modulation of Protein:Protein Interactions
NanoBiT® BiBiT PPI Starter System enables sensitive, quantitative measurement of HER2:HER3 interactions in response to therapeutic antibodies. Using stable cell lines expressing NanoBiT-tagged HER2 and HER3, differential dose responses to pertuzumab, trastuzumab and rituximab were observed. These results illustrate how NanoBiT® PPI assays can distinguish functional antibody effects on receptor engagement, making it an ideal platform for screening and characterizing therapeutic candidates in a physiologically relevant context.
Choose Starter Systems, Prebuilt or Custom NanoBiT® Assays
Get started with NanoBiT® PPI Assays in a format that suits your needs. Design your own assays using the NanoBiT® Starter Systems, or choose from prebuilt, optimized assays for a number of popular research targets including our newest collection of RAS pathway interaction assays. You can consult with us to design a customized assay specific to your research.
Protocols
No protocols available
Specifications
Catalog Number:
Promega DNA Vector Limited Use Label License (LULL)
BY USE OF THIS MATERIAL, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
-
1. Usage Restrictions:
- 1.1. Recipient may use this material for research use only; no commercial use of this material is allowed without the written consent of Promega Corporation.
- 1.2. “Commercial use” means any and all uses of this material or derivatives by recipient for monetary or other consideration, including but not limited to product manufacture or resale of the material for any use and sale of derivatives for any use.
- 1.3. If recipient is a contract research organization (“CRO”), recipient may transfer or sell data or information generated using the material or derivatives to CRO’s customers who request such data or information, provided that the material or derivatives cannot be transferred from CRO to CRO’s customers.
-
2. Modification Restrictions:
- 2.1. Recipient shall have no right to modify or otherwise create variations of the nucleotide sequence of any proprietary gene including but not limited to luciferase, NanoBiT® technology (e.g., HiBiT), HaloTag® technology or genes included in fusion vectors.
- 2.2. Recipient shall have the right to make derivatives of cloning vectors by incorporating exogenous sequences provided the resulting fusion gene has <4 nucleotides deleted from the affected terminus of the proprietary genes.
-
3. For determinations of luminescence activity of this material and its derivatives, recipient must either:
- 3.1. Use Promega-branded luminescent assay reagents (LARs); or
- 3.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
4. For determinations of NanoLuc® or NanoBiT® luminescence activity of this material and its derivatives, recipient must either:
- 4.1. Use Nano-Glo®-branded LARs; or
- 4.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
5. For determinations of GloSensor™ activity of this material and its derivatives, recipient must either:
- 5.1. Use GloSensor™ cAMP Reagent for in vitro and in-cell applications, and VivoGlo™ Luciferin, In Vivo Grade, for animal applications for all determinations of luminescence activity of this product and derivatives); or
- 5.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
6. For uses of this material for energy transfer (such as bioluminescence resonance energy transfer), recipient must:
- 6.1. Use NanoBRET™-branded luminescent assay reagents (LARs; e.g., NanoBRET™ Nano-Glo® Substrate), Intracellular TE Nano-Glo® Substrate/Inhibitor or Intracellular TE Nano-Glo® Vivazine™/Inhibitor for all determinations of luminescent activity by this material and its derivatives; and
- 6.2. Use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this material and its derivatives; or
- 6.3. Contact Promega to obtain a license for the use of the material and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded auto-fluorescent protein.
-
7. For uses of HaloTag® Technology in this material, recipient must either:
- 7.1. Use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties; or
- 7.2. Contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
-
8. Transfer of Materials:
- 8.1. Unmodified material or progeny of such material cannot be transferred.
- 8.2. Derivatives of cloning vectors can be transferred to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license.
- 8.3. If recipient is not a CRO, user may transfer this material to a CRO in order to have CRO use the material solely on behalf of recipient to provide recipient data or information, and CRO may transfer or sell such data or information to recipient, provided that recipient provides CRO with a copy of this label license and CRO agrees to comply with this label license.
- 8.4. Notwithstanding any other provision in this label license, the recipient can transfer the material (original or derivative of a cloning vector) to a third party for the sole purpose of amplification of the vector, provided that at the time of transfer a copy of this label license is given to the third party and the third party agrees to be bound by the terms of this label license. The third party may not use any of the material (original or amplified) for its own purposes or further transfer the material to any other party.
-
9. Other Uses:
- 9.1. For any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, contact Promega for supply and licensing information.
-
10. Disclaimer and Governing Law:
- 10.1. 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 MATERIAL. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
Promega DNA Vector Limited Use Label License (LULL)
BY USE OF THIS MATERIAL, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
-
1. Usage Restrictions:
- 1.1. Recipient may use this material for research use only; no commercial use of this material is allowed without the written consent of Promega Corporation.
- 1.2. “Commercial use” means any and all uses of this material or derivatives by recipient for monetary or other consideration, including but not limited to product manufacture or resale of the material for any use and sale of derivatives for any use.
- 1.3. If recipient is a contract research organization (“CRO”), recipient may transfer or sell data or information generated using the material or derivatives to CRO’s customers who request such data or information, provided that the material or derivatives cannot be transferred from CRO to CRO’s customers.
-
2. Modification Restrictions:
- 2.1. Recipient shall have no right to modify or otherwise create variations of the nucleotide sequence of any proprietary gene including but not limited to luciferase, NanoBiT® technology (e.g., HiBiT), HaloTag® technology or genes included in fusion vectors.
- 2.2. Recipient shall have the right to make derivatives of cloning vectors by incorporating exogenous sequences provided the resulting fusion gene has <4 nucleotides deleted from the affected terminus of the proprietary genes.
-
3. For determinations of luminescence activity of this material and its derivatives, recipient must either:
- 3.1. Use Promega-branded luminescent assay reagents (LARs); or
- 3.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
4. For determinations of NanoLuc® or NanoBiT® luminescence activity of this material and its derivatives, recipient must either:
- 4.1. Use Nano-Glo®-branded LARs; or
- 4.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
5. For determinations of GloSensor™ activity of this material and its derivatives, recipient must either:
- 5.1. Use GloSensor™ cAMP Reagent for in vitro and in-cell applications, and VivoGlo™ Luciferin, In Vivo Grade, for animal applications for all determinations of luminescence activity of this product and derivatives); or
- 5.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
6. For uses of this material for energy transfer (such as bioluminescence resonance energy transfer), recipient must:
- 6.1. Use NanoBRET™-branded luminescent assay reagents (LARs; e.g., NanoBRET™ Nano-Glo® Substrate), Intracellular TE Nano-Glo® Substrate/Inhibitor or Intracellular TE Nano-Glo® Vivazine™/Inhibitor for all determinations of luminescent activity by this material and its derivatives; and
- 6.2. Use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this material and its derivatives; or
- 6.3. Contact Promega to obtain a license for the use of the material and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded auto-fluorescent protein.
-
7. For uses of HaloTag® Technology in this material, recipient must either:
- 7.1. Use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties; or
- 7.2. Contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
-
8. Transfer of Materials:
- 8.1. Unmodified material or progeny of such material cannot be transferred.
- 8.2. Derivatives of cloning vectors can be transferred to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license.
- 8.3. If recipient is not a CRO, user may transfer this material to a CRO in order to have CRO use the material solely on behalf of recipient to provide recipient data or information, and CRO may transfer or sell such data or information to recipient, provided that recipient provides CRO with a copy of this label license and CRO agrees to comply with this label license.
- 8.4. Notwithstanding any other provision in this label license, the recipient can transfer the material (original or derivative of a cloning vector) to a third party for the sole purpose of amplification of the vector, provided that at the time of transfer a copy of this label license is given to the third party and the third party agrees to be bound by the terms of this label license. The third party may not use any of the material (original or amplified) for its own purposes or further transfer the material to any other party.
-
9. Other Uses:
- 9.1. For any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, contact Promega for supply and licensing information.
-
10. Disclaimer and Governing Law:
- 10.1. 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 MATERIAL. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
Promega DNA Vector Limited Use Label License (LULL)
BY USE OF THIS MATERIAL, RECIPIENT AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE.
-
1. Usage Restrictions:
- 1.1. Recipient may use this material for research use only; no commercial use of this material is allowed without the written consent of Promega Corporation.
- 1.2. “Commercial use” means any and all uses of this material or derivatives by recipient for monetary or other consideration, including but not limited to product manufacture or resale of the material for any use and sale of derivatives for any use.
- 1.3. If recipient is a contract research organization (“CRO”), recipient may transfer or sell data or information generated using the material or derivatives to CRO’s customers who request such data or information, provided that the material or derivatives cannot be transferred from CRO to CRO’s customers.
-
2. Modification Restrictions:
- 2.1. Recipient shall have no right to modify or otherwise create variations of the nucleotide sequence of any proprietary gene including but not limited to luciferase, NanoBiT® technology (e.g., HiBiT), HaloTag® technology or genes included in fusion vectors.
- 2.2. Recipient shall have the right to make derivatives of cloning vectors by incorporating exogenous sequences provided the resulting fusion gene has <4 nucleotides deleted from the affected terminus of the proprietary genes.
-
3. For determinations of luminescence activity of this material and its derivatives, recipient must either:
- 3.1. Use Promega-branded luminescent assay reagents (LARs); or
- 3.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
4. For determinations of NanoLuc® or NanoBiT® luminescence activity of this material and its derivatives, recipient must either:
- 4.1. Use Nano-Glo®-branded LARs; or
- 4.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
5. For determinations of GloSensor™ activity of this material and its derivatives, recipient must either:
- 5.1. Use GloSensor™ cAMP Reagent for in vitro and in-cell applications, and VivoGlo™ Luciferin, In Vivo Grade, for animal applications for all determinations of luminescence activity of this product and derivatives); or
- 5.2. Contact Promega to obtain a license for the use of LARs not manufactured by Promega.
-
6. For uses of this material for energy transfer (such as bioluminescence resonance energy transfer), recipient must:
- 6.1. Use NanoBRET™-branded luminescent assay reagents (LARs; e.g., NanoBRET™ Nano-Glo® Substrate), Intracellular TE Nano-Glo® Substrate/Inhibitor or Intracellular TE Nano-Glo® Vivazine™/Inhibitor for all determinations of luminescent activity by this material and its derivatives; and
- 6.2. Use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this material and its derivatives; or
- 6.3. Contact Promega to obtain a license for the use of the material and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded auto-fluorescent protein.
-
7. For uses of HaloTag® Technology in this material, recipient must either:
- 7.1. Use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties; or
- 7.2. Contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
-
8. Transfer of Materials:
- 8.1. Unmodified material or progeny of such material cannot be transferred.
- 8.2. Derivatives of cloning vectors can be transferred to others for research use provided that at the time of transfer a copy of this label license is given to the recipients and recipients agree to be bound by the terms of this label license.
- 8.3. If recipient is not a CRO, user may transfer this material to a CRO in order to have CRO use the material solely on behalf of recipient to provide recipient data or information, and CRO may transfer or sell such data or information to recipient, provided that recipient provides CRO with a copy of this label license and CRO agrees to comply with this label license.
- 8.4. Notwithstanding any other provision in this label license, the recipient can transfer the material (original or derivative of a cloning vector) to a third party for the sole purpose of amplification of the vector, provided that at the time of transfer a copy of this label license is given to the third party and the third party agrees to be bound by the terms of this label license. The third party may not use any of the material (original or amplified) for its own purposes or further transfer the material to any other party.
-
9. Other Uses:
- 9.1. For any uses outside this label license, including any diagnostic, therapeutic, prophylactic or commercial uses, contact Promega for supply and licensing information.
-
10. Disclaimer and Governing Law:
- 10.1. 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 MATERIAL. The terms of this label license shall be governed under the laws of the State of Wisconsin, USA.
Resources
Featured Resource
Proven BiBiT Assay Performance
For a detailed demonstration of the BiBiT approach in stable cell line development, refer to our poster, which outlines the use of BiBiT-ready vectors for stable, balanced expression of LgBiT and SmBiT fusion proteins from a single genomic locus.
Highlighted examples include key membrane protein interactions such as HER2:HER3, VEGFR1:VEGFR1, CX3CR1:ARRB2 and CRAF:BRAF. This robust approach will maximize reproducibility and sensitivity in your assays.
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NanoBiT® PPI Control Vectors
Validated positive, negative and performance controls for NanoBiT® PPI assay workflows.
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FuGENE® HD Transfection Reagent
Enhanced performance in a range of cell types, including difficult-to-transfect cell lines.
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FuGENE® 4K Transfection Reagent
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