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NanoLuc® Stability Sensors for Cell Signaling

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Vectors to Measure Intracellular Protein Turnover of HIFA and NRF2

  • Ready-to-use constructs are predesigned, optimized and tested for low endotoxin levels
  • C-terminal fusion of HIF1A NanoLuc® luciferase reporter in pNLF1-HIF1A [CMV/neo] Vector
  • NRF2 vector system has a pKEAP1-expressing vector for regulating intracellular NRF2 levels and pNLF1-NRF2 [CMV/neo] Vector expressing a C-terminal fusion with NanoLuc® luciferase

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Catalog number selected: N1381

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NanoLuc® Stability Sensors for Cell Signaling
HIF1A/20µg
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The rate of protein turnover is tightly regulated for many signaling proteins involved in oncogenesis and response to cellular stress. Protein stabilization and subsequent accumulation occurs in response to changing cellular conditions resulting in activation of downstream transcriptional events. The NanoLuc® Stability Sensors are ready-to-use vector systems that utilize the advantages of the NanoLuc® luciferase reporter to enable stability studies of two key signaling proteins, HIF1A and NRF2, providing a method to directly measure this primary signaling event.

HIF1A Vector System: The HIF1A Vector System enables simple quantification of intracellular HIF1A protein levels to study the dynamics of this signaling protein in mediating cellular response to hypoxia. It contains a vector encoding NanoLuc® fused to the C-terminus of the HIF1A protein under control of the CMV promoter plus Transfection Carrier DNA for titrating intracellular fusion protein expression.

NRF2 Vector System: The NRF2 Vector System enables simple quantification of intracellular NRF2 protein levels to study the dynamics of this signaling protein in mediating cellular response to oxidative stress. It contains a vector encoding NanoLuc® fused to the C-terminus of the NRF2 protein under the control of the CMV promoter, a pKEAP1-expressing vector for proper regulation of intracellular NRF2 levels and Transfection Carrier DNA for titratable intracellular fusion protein expression.

Applications

  • Compound screening.
  • Hypoxia signaling.
  • Oxidative stress signaling.

Sequence Information

pNLF1-HIF1A [CMV/neo] Vector GenBank® Accession Number KF853601 and vector sequence text file.

pNLF1-NRF2 [CMV/neo] Vector GenBank® Accession Number KF853602 and vector sequence text file
(pKEAP1 [CMV/Hygro] Vector GenBank® Accession Number KF853604 and vector sequence text file).

Rapid stabilization and accumulation of NRF2 monitored with NRF2-NanoLuc® fusion protein.

Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.

Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.

Rapid stabilization and accumulation of HIF1A monitored with HIF1A-NanoLuc® fusion protein.

Specifications

You are viewing: N1381 Change Configuration

What's in the box?

Item Part # Size Concentration Available Separately

Transfection Carrier DNA

E488A 1 × 20μg 1μg/μl View Product

pNLF1-HIF1A [CMV/neo] Vector

N138A 1 × 20μg 1μg/μl

Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

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.

Specifications

You are viewing: N1391 Change Configuration

What's in the box?

Item Part # Size Concentration Available Separately

Transfection Carrier DNA

E488A 1 × 20μg 1μg/μl View Product

pNLF1-NRF2 [CMV/neo] Vector

N139A 1 × 20μg 1μg/μl

pKEAP1[CMV/Hygro] Vector

N140A 1 × 20μg 1μg/μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

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.

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