N1870_NanoBRET--cMyc-3aMAX-Interaction-Asy_3

Sensitive Live-Cell Assays for Monitoring Proteins Interactions in Transcription Regulation

  • Order NanoBRET® cMyc/MAX Interaction Assay by selecting N1870
  • For additional transcriptional assays go to "View Prebuilt Assays" link below

Catalog Number:

Size

Catalog Number: N1870

Please Enquire


Overview
Protocols
Specifications
Resources
Related Products

細胞内環境で真のタンパク質相互作用研究を

NanoBRET™ Transcriptional Protein Assayは、転写調節に関与するタンパク質の相互作用をモニタリング/スクリーニングするためにデザインされた高感度で再現性の高い生細胞アッセイです。タンパク質間の相互作用は、DNAエレメントにホモダイマー/ヘテロダイマーが結合することで制御される遺伝子発現において主要な役割を果たしており、転写イベントの制御は様々な細胞応答に必要です。NanoBRET™ cMyc/MAX Interaction Assay は本来の細胞内環境でヒトcMycと Max転写因子との特異的な相互作用を測定します。cMyc/MAXヘテロダイマーは細胞の増殖、分化、アポトーシスに関連する転写を制御し、薬剤標的の重要な標的の候補として考えれれています。

NanoBRET™ assay テクノロジーは生物発光ドナー(NanoLuc® ルシフェラーゼ)から蛍光アクセプター(HaloTag® NanoBRET™ 618 Ligand)へのエネルギー転移をベースにしています。NanoLuc® ルシフェラーゼおよび HaloTag® タンパク質を標的となるタンパク質にそれぞれ融合させ、低レベルで細胞内に発現させます。これにより真の細胞生理を反映するタンパク質相互作用のモニタリングやスクリーニングの実験を行うことができます。 NanoBRET™ assay は完全な可逆性を有しており、タンパク質相互作用の誘導実験、阻害実験の両方を行うことができます。

Q&A NanoBRET 
【ライセンス】NanoBRET / NanoLuc / HaloTag

優れたドナーおよびアクセプターのシグナル分離度

Energy Transfer from NanoLuc to HaloTag NanoBRET Ligand 12749TA-W-a
Depiction of energy transfer from a NanoLuc®-Protein A fusion (energy donor) to a fluorescently labeled HaloTag®-Protein B fusion (energy acceptor) upon interaction of Protein A and Protein B. 
Donor and Acceptor Signal Wavelengths for NanoBRET
Spectral separation of the NanoLuc® emission (460nm) and the fluorescent HaloTag® NanoBRET® 618 Ligand emission (618nm), and calculation of the NanoBRET® ratio.

備 考

  • Sufficient NanoBRET™ Nano-Glo® Substrate and HaloTag® NanoBRET™ 618  Ligand are provided to perform 400 assays in 96-well plates or 1,000 assays in 384-well plates.
  • To perform NanoBRET™ PPI Assays an instrument, such as the GloMax® Discover, capable of sequentially measuring dual-filtered luminescence and equipped with appropriate filters is required. The ideal filter setup will include a band pass (BP) filter centered around 460nm to measure the donor signal (e.g., Emission 450nm/BP 80nm) and a long pass (LP) filter starting at around 600–610nm to measure the acceptor signal (e.g., Emission 610nm/LP). Filters outside of these ranges will miss critical measurements and compromise data quality.

Obtain High Assay Specificity

MAX-HaloTag and cMyc-NanoLuc DSA Specificity
Donor saturation assay performed by transfecting a constant amount of cMyc-NanoLuc® donor DNA and increasing amounts of MAX-HaloTag® acceptor DNA to represent increasing acceptor-to-donor (A:D) ratios. The negative control is nuclear-localized NanoLuc® protein as a mock donor in place of the cMyc-NanoLuc® protein fusion. The NanoBRET® cMyc/MAX Interaction Assay generates a hyperbolic curve, which indicates detection of a specific BRET interaction.

スターターセットおよび”構築済み” または ""カスタム" アッセイも利用可能

ご要望に合ったフォーマットの NanoBRET™ PPI Assay をお選びいただけます。スターターシステムならご自身でアッセイを構築できます。またプレデザインされたアッセイ(カタログ品)からお選びいただくこともカスタムアッセイを構築することも可能です。

Transcriptional Protein Assay Vectors
12745MA-W
cMyc-NanoLuc® Fusion Vector sequence text file.
12746MA-W
MAX-HaloTag® Fusion Vector sequence text file.
NanoBRET® Control Vectors
12729MA-W
NanoBRET® Positive Control Vector sequence text file.
12730MA-W
p53-HaloTag® Fusion Vector sequence text file.
12731MA-W
NanoLuc®-MDM2 Fusion Vector sequence text file.

Specifications

Catalog Number:

選択製品の構成品内容

Item Part # Size Available Separately

HaloTag® NanoBRET® 618 Ligand

G980A 2 × 20μl View Product

NanoBRET® Nano-Glo® Substrate

N157A 2 × 50μl View Product

p53-HaloTag® Fusion Vector

N159A 1 × 20μg

NanoLuc®-MDM2 Fusion Vector

N160A 1 × 20μg

cMyc-NanoLuc® Fusion Vector

N179A 1 × 20μg

MAX-HaloTag® Fusion Vector

N180A 1 × 20μg

分析証明書

Search by lot number

使用制限

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

保存条件

BB

パテントおよび免責条項

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 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. Researcher shall have no right to modify or otherwise create variations of the nucleotide sequence of the luciferase gene or the HaloTag® gene except that researcher may create fused gene sequences provided that the coding sequence of the resulting luciferase gene or HaloTag® gene has no more than four deoxynucleotides missing at the affected terminus compared to the intact gene sequence. No other use of this product or derivatives 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 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, researcher 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.

Researcher 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 label license shall be governed under the laws of the State of Wisconsin, USA.

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 commercial use 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 product. No other use or transfer of this product is authorized without the prior express written consent of Promega.

For uses of Nano-Glo®-branded reagents intended for energy transfer (such as bioluminescence resonance energy transfer) to acceptors other than a genetically encoded autofluorescent protein, researchers must:
(i) use NanoBRET®-branded energy acceptors (e.g., BRET-optimized HaloTag® ligands) for all determinations of energy transfer activity by this product; or
(ii) contact Promega to obtain a license for use of the product for energy transfer assays to energy acceptors not manufactured by Promega.

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. 8,420,367, 8,748,148, 9,593,316 and 9,873,866; European Pat. Nos. 2374875, 2492342 and 2502990; and Japanese Pat. Nos. 5680302 and 5840117.

U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.

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.

U.S. Pat. No. 10,024,862; Japanese Pat. No. 7676487; and other patents and patents pending.

U.S. Pat. Nos. 9,056,885, 10,139,400 and 10,215,751; European Pat. No. 2782916; and Japanese Pat. Nos. 6522339, 6574821 and 7227741.

Resources

No related resources available