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Bidirectional Vector for Validating NanoBRET® BiBRET Assays

  • Optimized BiBRET vector designed to measure the interaction of protein pairs CRAF and BRAF using NanoBRET® Nano-Glo® Detection Systems
  • Include as a reference control vector when performing other NanoBRET® BiBRET PPI assays

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Catalog Number: CS1679D24

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Single-Vector Control for BiBRET Assay Performance​

The CRAF/BRAF BiBRET Control Vector is a bidirectional expression plasmid designed to co-express HaloTag-CRAF and NanoLuc-BRAF fusion proteins from a single vector. This construct serves as a functional positive control for BiBRET PPI assays, enabling researchers to confirm assay performance and signal detection in live-cell environments. Using the same bidirectional CMV promoter architecture as the BiBRET PPI Starter System, this vector ensures consistent expression of both fusion partners for reproducible protein:protein interaction readouts and is ideal for assay optimization, troubleshooting and benchmarking across experiments.

CRAF:BRAF Control Response to GDC0879

 

 

 

HEK293 cells transiently transfected with the CRAF:BRAF BiBRET Control Vector displayed a clear, dose-dependent increase in BRET ratio following treatment with the RAF inhibitor GDC0879. NanoBRET® ratios were measured using NanoBRET® Nano-Glo® Detection System. The resulting EC50 of ~1.26μM demonstrates effective PPI induction and highlights the utility of this control vector for benchmarking BiBRET assay performance in live-cell systems.

Line graph showing a clear, dose-dependent increase in BRET ratio for HEK293 cells transiently transfected with the CRAF:BRAF Control Vector and treated with the RAF inhibitor GDC0879.

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Patents and Disclaimers

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.

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