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NanoBRET™ TE E3 Ligase Vectors

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NanoLuc® E3 Ligase Fusion Vectors and DDB1 Expression Vectors for NanoBRET™ TE E3 Ligase Assays

  • Transfection-ready vectors to express NanoLuc® luciferase fused to E3 ligase CRBN or VHL proteins
  • Transfection-ready vector to express DDB1 protein
  • Use these vectors with NanoBRET™ TE Intracellular E3 Ligase Assays, CRBN or VHL, to measure E3 ligase target engagement

Choose a Fusion or Expression Vector

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Catalog number selected: N2741

$ 515.00
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NanoBRET™ TE E3 Ligase Vectors
NanoLuc®-CRBN/20µg
$ 515.00
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Study Cellular E3 Ligase Target Engagement

Protein degradation via small molecules such as PROTACs or molecular glues is governed by multiple steps. The initial steps involve entry of the small molecule degrader compound into the cell, followed by binding to the E3 ubiquitin ligase and protein target. The NanoBRET™ Target Engagement (TE) Intracellular E3 Ligase Assays allow live-cell measurement of compound binding or affinity for an E3 ubiquitin ligase protein, Cereblon (CRBN) or von Hippel-Lindau disease tumor suppressor (VHL). In addition, the assays can be used to quantify intracellular availability of the compound. These assays are based on the NanoBRET™ technology, an energy transfer technique designed to measure molecular proximity in living cells.

The NanoLuc® E3 ligase fusion vectors are designed to be the energy donor in the NanoBRET™ TE Intracellular E3 Ligase Assay. The energy acceptor in these assays is a fluorescent cell-permeable NanoBRET™ Tracer that binds to the NanoLuc® E3 ligase fusion protein. The close proximity of the tracer to NanoLuc® in the E3 ligase fusion results in a NanoBRET™ signal. Addition of compounds to cells that bind the E3 ligase results in a dose-dependent decrease in NanoBRET™ signal. This data allows quantitation of the intracellular affinity of degrader compounds for a target E3 ligase protein.

NanoLuc®-CRBN Fusion Vector

The NanoLuc®-CRBN Fusion Vector contains the coding region of human E3 ligase component cereblon (CRBN) fused to the C-terminus of the NanoLuc® luciferase coding region. The vector can be transfected into various cell lines for CRBN target engagement analysis using the NanoBRET™ Target Engagement (TE) Intracellular E3 Ligase Assay CRBN. In this assay, it is recommended that the NanoLuc®-CRBN Fusion Vector be used with the DDB1 Expression Vector (N2761). The NanoLuc®-CRBN Fusion Vector is supplied transfection-ready. The CMV promoter drives the expression of the full-length NanoLuc®-CRBN fusion protein. For more information please see the NanoBRET™ TE Intracellular E3 Ligase Assays.

NanoLuc-CRBN Fusion Vector

NanoLuc®-CRBN Fusion Vector sequence text file.

VHL-NanoLuc® Fusion Vector

The VHL-NanoLuc® Fusion Vector contains the coding region of human von Hippel-Lindau disease tumor suppressor E3 ligase (VHL) fused to the N-terminus of the NanoLuc® luciferase coding region. The vector can be transfected into various cell lines for VHL target engagement analysis using the NanoBRET™ Target Engagement (TE) Intracellular E3 Ligase Assay VHL. The VHL-NanoLuc® Fusion Vector is supplied transfection-ready. The CMV promoter drives the expression of the full-length VHL-NanoLuc® fusion protein. For more information please see the NanoBRET™ TE Intracellular E3 Ligase Assays.

VHL-NanoLuc Fusion Vector

VHL-NanoLuc® Fusion Vector sequence text file.

DDB1 Expression Vector

The DDB1 Expression Vector contains the coding region of human DNA damage-binding protein 1 (DDB1), a component of a E3 ubiquitin ligase complex. The DDB1 Expression Vector is designed for use with the NanoLuc®-CRBN Fusion Vector (N2741) and the NanoBRET™ TE Intracellular E3 Ligase Assay CRBN. The DDB1 Expression Vector is supplied transfection-ready. The CMV promoter drives the expression of the full-length DDB1 protein. For more information please see the NanoBRET™ TE Intracellular E3 Ligase Assays.

DBB1 Expression Vector

DDB1 Expression Vector sequence text file.

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Specifications

You are viewing: N2741 Change Configuration

What's in the box?

Item Part # SizeConcentration

NanoLuc®-CRBN Fusion Vector

N274A 1 × 20μg1μg/μl

SDS

Choose language:

Download SDSPDF (191 KB) – English (United States)

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

NanoLuc® ORF TE Vector Limited Use Label License

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 terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.

Researcher 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 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, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene. No other use of this product is authorized without the prior express written consent of Promega.

In addition, researcher must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product; or
(1b) contact Promega to obtain a license for use of the product with LARs not manufactured by Promega.

For uses of this product (NanoLuc® ORF Vector) for energy transfer (such as bioluminescence resonance energy transfer), researcher must:
(2a) 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; and
(2b) use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity; or
(2c) contact Promega to obtain a license for use of the product for energy transfer assays using energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.

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 label license 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 and 10,633,690 and other patents and patents pending.

Licensed from Kazusa Genome Technologies.

Specifications

You are viewing: N2751 Change Configuration

What's in the box?

Item Part # SizeConcentration

VHL-NanoLuc® Fusion Vector

N275A 1 × 20μg1μg/μl

SDS

Choose language:

Download SDSPDF (190 KB) – English (United States)

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

NanoLuc® ORF TE Vector Limited Use Label License

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 terms of this label license, and the product is unused, Promega will accept return of the unused product and provide researcher with a full refund.

Researcher 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 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, whether or not such product is resold for use in research. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene. No other use of this product is authorized without the prior express written consent of Promega.

In addition, researcher must:
(1a) use Nano-Glo®-branded luminescent assay reagents (LARs) for all determinations of luminescence activity of this product; or
(1b) contact Promega to obtain a license for use of the product with LARs not manufactured by Promega.

For uses of this product (NanoLuc® ORF Vector) for energy transfer (such as bioluminescence resonance energy transfer), researcher must:
(2a) 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; and
(2b) use NanoBRET™-branded energy acceptors (e.g., NanoBRET™ tracers, NanoBRET™ dyes, BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity; or
(2c) contact Promega to obtain a license for use of the product for energy transfer assays using energy acceptors not manufactured by Promega. No license is needed if the energy transfer acceptor is a genetically encoded autofluorescent protein.

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 label license 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 and 10,633,690 and other patents and patents pending.

Licensed from Kazusa Genome Technologies.

Specifications

You are viewing: N2761 Change Configuration

What's in the box?

Item Part # SizeConcentration

DDB1 Expression Vector

N276A 3 × 20μg1μg/μl

SDS

Choose language:

Download SDSPDF (189 KB) – English (United States)

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

Licensed from Kazusa Genome Technologies.

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