pNLF1 NanoLuc® Protein Fusion Vectors
Generate N- or C-Terminal Fusions with NanoLuc® Enzyme or N-Terminal Secreted NanoLuc® Luciferase
- Use traditional cloning to insert protein of interest into the multiple cloning site (MCS)
- Bright NanoLuc® reporter enables endogenous expression levels of fusion proteins for biologically relevant results
- Choose from transient transfection or select for stable expression using hygromycin
Catalog Number:
Choose a protein fusion
Size
Catalog Number: N1351
Catalog Number: N1361
Catalog Number: N1371
The pNLF1 Vectors use traditional cloning with a multiple cloning site (MCS) to generate N- or C-terminal fusions to NanoLuc® luciferase. Create full-length Nluc protein fusions using the pNLF1-N [CMV/Hygro] Vector (N terminus) and pNLF1-C [CMV/Hygro] Vector (C terminus). In addition, attach secreted Nluc to the N terminus of the protein of interest using the pNLF1-secN [CMV/Hygro] Vector. All vectors contain a mammalian selectable marker to create a stable line.
The small size (19.1kDa) and extreme brightness of NanoLuc® luciferase (Nluc; about 100-fold brighter than either firefly [Photinus pyralis] or Renilla reniformis) make it an ideal protein fusion partner. When visualizing intracellular protein dynamics, the bright NanoLuc® reporter reduces imaging exposure times without the need for repeated sample excitation, which can result in cytotoxic artifacts.
NanoLuc® fusion proteins can be used in a variety of applications including: reporters of protein stability, probes for bioluminescent cell imaging (BLI) or as the donor signal in bioluminescent resonance energy transfer (BRET) applications for protein:protein or protein:small-molecule interaction studies. The brighter signal and blue-shifted emission spectrum from NanoLuc® luciferase result in less spectral overlap with fluorescent acceptors, resulting in better signal:background and dynamic range for BRET applications.
Additional NanoLuc® protein fusion vectors include the pFN31 and pFC32 Vectors. Generate N- or C-terminal Nluc fusion proteins using the Flexi® Vector Cloning System—a directional cloning method based on two rare-cutting restriction enzymes, SgfI and PmeI, that provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi® Vectors without the need to resequence.
Applications
- Compound screening.
- Protein:protein interactions.
- Protein:small-molecule interactions.
- Bioluminescent imaging.
- Protein stability.
Reference
- Hall, M.P. (2012) Engineered luciferase reporter from a deep sea shrimp utilizing a novel imidazopyrazinone substrate. ACS Chemical Biology 7(11), 1848–57.
pNLF1-N [CMV/Hygro] Vector GenBank® Accession Number KF811457 and vector sequence text file.
pNLF1-C [CMV/Hygro] Vector GenBank® Accession Number KF811458 and vector sequence text file.
pNLF1-secN [CMV/Hygro] Vector GenBank® Accession Number KF811459 and vector sequence text file.
Protocols
No protocols available
Specifications
Catalog Number:
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
pNLF1-N [CMV/Hygro] Vector |
N135A | 1 × 20μg | 1μg/μl |
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 conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researchers may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product or derivatives 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 or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that researcher may (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence; and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use of this product or derivatives is authorized without the prior express written consent of Promega.
In addition, researchers must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product and its derivatives; or
(1b) contact Promega to obtain a license for use of the product and its derivatives with LARs not manufactured by Promega.
For uses of Nano-Glo®-branded LARs intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(2a) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product and its derivatives; or
(2b) contact Promega to obtain a license for use of the product and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega.
Researchers may transfer derivatives 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. 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 agreement shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
pNLF1-C [CMV/Hygro] Vector |
N136A | 1 × 20μg | 1μg/μl |
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 conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researchers may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product or derivatives 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 or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that researcher may (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence; and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use of this product or derivatives is authorized without the prior express written consent of Promega.
In addition, researchers must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product and its derivatives; or
(1b) contact Promega to obtain a license for use of the product and its derivatives with LARs not manufactured by Promega.
For uses of Nano-Glo®-branded LARs intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(2a) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product and its derivatives; or
(2b) contact Promega to obtain a license for use of the product and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega.
Researchers may transfer derivatives 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. 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 agreement shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
Contenido
| Item | Part # | Presentación | Concentración |
|---|---|---|---|
pNLF1-secN [CMV/Hygro] Vector |
N137A | 1 × 20μg | 1μg/μl |
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 conditions of this limited use statement, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.
Researchers may use this product for research use only; no transfer or commercial use of this product is allowed. Commercial use means any and all uses of this product or derivatives 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 or its derivatives, whether or not such product or derivatives are resold for use in research. Researchers shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene except that researcher may (1) create fused gene sequences provided that the coding sequence of the resulting luciferase gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact luciferase gene sequence; and (2) insert and remove nucleic acid sequences in splicing research predicated on the inactivation or reconstitution of the luminescence of the encoded luciferase. No other use of this product or derivatives is authorized without the prior express written consent of Promega.
In addition, researchers must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product and its derivatives; or
(1b) contact Promega to obtain a license for use of the product and its derivatives with LARs not manufactured by Promega.
For uses of Nano-Glo®-branded LARs intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(2a) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product and its derivatives; or
(2b) contact Promega to obtain a license for use of the product and its derivatives for energy transfer assays to energy acceptors not manufactured by Promega.
Researchers may transfer derivatives 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. 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 agreement shall be governed under the laws of the State of Wisconsin, USA.
U.S. Pat. Nos. 8,557,970, 8,669,103, 9,777,311, 9,840,730, 9,951,373, 10,233,485, 10,633,690, 10,774,364, 10,844,422, 11,365,436, 11,661,623, 11,667,950; European Pat. Nos. 2456864, 2635595, 2990478, 3181687, 3409764; Japanese Pat. Nos. 6038649, 6155424, 6227615, 6374420, 6539689; and other patents and patents pending.
Resources
No related resources available
Related Products
Productos Similares
pFN31 and pFC32 NanoLuc® Protein Fusion Flexi® Vectors
Cuatro vectores Flexi® Cloning de fusión de proteínas para generar fusiones N- o C-terminal de la NanoLuc® Luciferase con su proteína de interés.
N1311, N1321, N1331, N1341
Transfection Carrier DNA
ADN plasmídico utilizado para reducir la cantidad de un vector de expresión o vector reportero en la transfección de células de mamífero, manteniendo la cantidad total de ADN.
E4881, E4882
NanoLuc® Stability Sensors for Cell Signaling
Sistemas de vectores que fusionan dos proteínas de señalización clave, HIF1A y NRF2, con la NanoLuc® Luciferasa para estudiar la estabilidad de la proteína de fusión.
N1381, N1391
Coincidence Reporter Vectors
Cuatro vectores con y sin promotores que expresan luciferasas firefly y NanoLuc® a partir del mismo transcrito para identificar falsos positivos durante el cribado.
N1461, N1471, N1481, N1491
Usado con frecuencia junto con
Nano-Glo® Luciferase Assay System
Ensayo "add-mix-measure" de NanoLuc® Luciferase con >2 horas de vida media en la mayoría de las aplicaciones.
N1110, N1120, N1130, N1150
FuGENE® HD Transfection Reagent
Rendimiento mejorado en una amplia gama de tipos celulares, incluidas líneas celulares difíciles de transfectar.
E2311, E2312
GloMax® Discover System
Lector de microplacas de alto rendimiento para la detección de luminiscencia, fluorescencia y absorbancia.
GM3000
