NanoBiT® PPI Starter Systems

Part Numbers: N2014, N2015, N2016

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生きた細胞で高感度なタンパク質相互作用検出

  • 発現レベルが低くてもPPI検出でき、生細胞でリアルタイムカイネティックアッセイも可能
  • 相補反応は可逆的であり、タンパク質の会合・解離の解析が可能
  • スプリットホタルルシフェラーゼを凌ぐ感度と正確性

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選択製品のカタログ番号: N2014

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NanoBiT® PPI Starter Systems
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細胞内で起こる真のタンパク質相互作用を測定

NanoLuc® Binary Technology (NanoBiT) はタンパク質:タンパク質相互作用 (PPI) 研究のためにデザインされた構造相補性レポーターです。NanoBiT® system は2つの小さなサブユニットである Large BiT (LgBiT; 18kDa) および Small BiT (SmBiT; 11 アミノ酸のペプチド)で構成されており、目的となるタンパク質ペアとそれぞれ融合タンパク質として発現させます。LgBiT および SmBiT の両サブユニットは安定性と互いの親和性が最小限になるよう最適化されています。これらのサブユニットをそれぞれ融合させた2つの目的タンパク質が相互作用すると、サブユニットも会合して1つの活性化酵素を形成し、これが明るい発光シグナルを生じます。

この NanoBiT 相補性パートナーが非常に小さいためノーマルなタンパク質機能への影響が最小限に抑えられ、さらに形成された酵素による非常に明るい光により通常の発現レベルでも十分にタンパク質相互作用を検出することができます。 

 NanoBiT アッセイのための コンストラクト作成のご相談 も承っております。

【ライセンス】プロメガライセンスプログラム

Study Drug Potency and Binding Kinetics with NanoBiT

薬効と結合カイネティクスの研究に

NanoBiTより得られる活性に比例したシグナルとプレートベースのフォーマットはハイスループットスクリーニグなどを含むタンパク質間相互作用検出に理想的です。

”NanoBiT は薬物作用の基本となるタンパク質相互作用の誘導とその結合カイネティクスに基づく薬効を明らかにすることができます。”

NanoBiT テクノロジーの基本的な技術情報とタンパク質相互作用アッセイのデータ例を以下の文献よりご覧いただけます。:NanoLuc Complementation Reporter Optimized for Accurate Measurement of Protein Interactions in Cells,  Dixon et al., (2016) ACS Chem. Biol., 11, 400–408.

文献を読む

タンパク質相互作用の誘導と阻害をモニタリングできるダイナミックで可逆的なアッセイ

多くのスプリットシステムと異なり、LgBiT:SmBiT 相互作用は可逆的であるためタンパク質の迅速な解離も検出することができます。細胞透過性のフリマジン基質を含む非細胞溶解性の検出試薬 Nano-Glo® Live Cell Reagent を用いることにより生細胞でのリアルタイムなタンパク質間相互作用ダイナミクスを追跡することができます。スプリットシステムに対して感度、可逆性などの点で優れており、ペプチドあるいは低分子で構造的に安定なタンパク質ドメインとの融合タンパク質を作成でき、タンパク質相互作用をリアルタイムで測定できることも有利な特性です。

技術ポスター ”Monitoring Intracellular Protein Interactions using NanoBiT Binary Technology” でこれらの特性に関する詳細なデーターをご覧いただけます。

ポスターをダウンロード
Monitor Real-Time PPI Dynamics in Cells with NanoBiT

リアルタイムにタンパク質ダイナミクスをモニタリング

この実験ではHeLa に FKBP-SmBiT および FRB-LgBiT の融合コンストラクトをトランスフェクションし、オーバーナイトでインキュベーション後、基質を加えてLV200 顕微鏡でサンプルのイメージングを行いました。約30秒間ベースラインを読んだ後に 500nM ラパマイシンを添加してFKBP:FRB 相互作用を誘導し、イメージを録画しました (イメージサイクル = 0.5 秒 )。

NanoBiT のアプリケーション例

NanoBiT® システムベクター

NanoBiT® PPI Starter System には LgBiT または SmBiT とのタンパク質 融合に必要なベクターおよびコントロールベクターならびに生細胞でNanoBiT®発光をモニタリングするためのワンショット - 非細胞溶解性の検出試薬 Nano-Glo® Live Cell Assay Systemが含まれます。各システムで供給されるベクターの詳細については以下をご覧ください。
NanoBiT® PPI MCS Starter System Vectors, Cat.# N2014

Using the NanoBiT® MCS Starter System, you can generate N- and C-terminal LgBiT and SmBiT fusions to proteins of interest using traditional cloning with a multiple cloning site (MCS). Expression is driven by HSV-TK promoter, providing constitutive, low-level expression in mammalian cells. 

13374MA-W
pBiT1.1-N[TK/LgBiT] Vector sequence text file
13373MA-W
pBiT2.1-C[TK/SmBiT] Vector sequence text file.
13369MA-W
SmBiT-PRKACA Control Vector sequence text file.
13372MA-W
pBiT1.1-C[TK/LgBiT] Vector sequence text file.
13367MA-W
NanoBiT® Negative Control Vector sequence text file.
13375MA-W
pBiT2.1-N[TK/SmBiT] Vector sequence text file.
13368MA-W
LgBiT-PRKAR2A Control Vector sequence text file.
NanoBiT® PPI Flexi® Starter System Vectors, Cat.# N2015

The NanoBiT® Flexi® Starter System, allows you to generate N- and C-terminal LgBiT and SmBiT fusions using the Flexi® Vector Cloning System, a directional cloning method based on the rare-cutting restriction enzymes, SgfI and PmeI, that provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between Flexi® Vectors without the need to resequence. The Find My Gene™ service contains many ORF clones already in Flexi® format for simple creation of fusions.

13370MA-W
pFN33K LgBiT TK-neo Flexi® Vector sequence text file.
13364MA-W
pFC36K SmBiT TK-Neo Flexi® Vector sequence text file.
13363MA-W
pFC34K LgBiT TK-Neo Flexi® Vector sequence text file.
13367MA-W
NanoBiT® Negative Control Vector sequence text file.
13371MA-W
pFN35K SmBiT TK-neo Flexi® Vector sequence text file.
13368MA-W
LgBiT-PRKAR2A Control Vector sequence text file.
13369MA-W
SmBiT-PRKACA Control Vector sequence text file.
pF4A CMV Flexi Vector.
pF4A CMV Flexi® Vector Accession Number AY753580; sequence text file.
NanoBiT® PPI Control  Pair (FKPB, FRB), Cat.# N2016

The NanoBiT® PPI Control Pair (FKBP:FRB; Cat.# N2016) is available separately as an inducible positive control.  

13365MA-W
FRB-LgBiT Control Vector sequence text file.
13366MA-W
FKBP-SmBiT Control Vector sequence text file.
Additional NanoBiT® Vectors and Assays
Alternative CMV-based Starter Systems compatible with stable cell line generation, additional control options, βarrestin recruitment vectors and optimized assays for hot targets such as KRAS and EGFR.

ニーズに合わせて選べるスターターシステム、既製アッセイ、カスタム設計

NanoBiT® PPI アッセイは、目的や研究スタイルに応じた形式で導入できます。NanoBiT® Starter System を使って独自のアッセイを設計することも、最適化された既製のアッセイを選ぶことも可能です。既製のアッセイには、最新のRAS経路に関する相互作用アッセイを含む、人気の研究ターゲットが多数揃っています。研究内容に合わせて、カスタムアッセイの設計についてご相談いただくことも可能です。

既成のアッセイを見る カスタムアッセイのお問合せ

製品仕様

You are viewing: N2014 Change Configuration

選択製品の構成品内容

Item Part # Size

pBiT1.1-C [TK/LgBiT] Vector

N196A 1 × 20μg

pBiT2.1-C [TK/SmBiT] Vector

N197A 1 × 20μg

pBiT1.1-N [TK/LgBiT] Vector

N198A 1 × 20μg

pBiT2.1-N [TK/SmBiT] Vector

N199A 1 × 20μg

NanoBiT® Negative Control Vector

N202A 1 × 20μg

LgBiT-PRKAR2A Control Vector

N203A 1 × 20μg

SmBiT-PRKACA Control Vector

N204A 1 × 20μg

Nano-Glo® Live Cell Substrate

N205A 2 × 125μl

Nano-Glo® LCS Dilution Buffer

N206A 2 × 2.5ml

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

See Protocol for detailed storage recommendations.

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.

No transfer of this product is allowed. Researchers may use this product in their own research and they may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to recipients and recipients agree to be bound by the terms of this label license.

Researchers shall have no right to modify or otherwise create variations of the nucleotide sequences of the subunits of NanoBiT® Technology, except that researchers may create fused gene sequences at the termini of the subunits of NanoBiT® Technology. No other use of this product or its derivatives is authorized without the prior express written consent of Promega.

In addition, researchers must either:
(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 either:
(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 has no right to modify, derivatize, genetically engineer or otherwise create variations of the NanoBiT® Control Vectors, except that researcher may propagate and store NanoBiT® Control Vectors for use in NanoBiT® assays. 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. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.

U.S. Pat. Nos. 9,797,889, 9,797,890 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents pending.

製品仕様

You are viewing: N2015 Change Configuration

選択製品の構成品内容

Item Part # Size

pF4A CMV Flexi® Vector

C848A 1 × 20μg

pFC34K LgBiT TK-neo Flexi® Vector

N192A 1 × 20μg

pFC36K SmBiT TK-neo Flexi® Vector

N193A 1 × 20μg

pFN33K LgBiT TK-neo Flexi® Vector

N194A 1 × 20μg

pFN35K SmBiT TK-neo Flexi® Vector

N195A 1 × 20μg

NanoBiT® Negative Control Vector

N202A 1 × 20μg

LgBiT-PRKAR2A Control Vector

N203A 1 × 20μg

SmBiT-PRKACA Control Vector

N204A 1 × 20μg

Nano-Glo® Live Cell Substrate

N205A 2 × 125μl

Nano-Glo® LCS Dilution Buffer

N206A 2 × 2.5ml

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

See Protocol for detailed storage recommendations.

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.

No transfer of this product is allowed. Researchers may use this product in their own research and they may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to recipients and recipients agree to be bound by the terms of this label license.

Researchers shall have no right to modify or otherwise create variations of the nucleotide sequences of the subunits of NanoBiT® Technology, except that researchers may create fused gene sequences at the termini of the subunits of NanoBiT® Technology. No other use of this product or its derivatives is authorized without the prior express written consent of Promega.

In addition, researchers must either:
(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 either:
(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 has no right to modify, derivatize, genetically engineer or otherwise create variations of the NanoBiT® Control Vectors, except that researcher may propagate and store NanoBiT® Control Vectors for use in NanoBiT® assays. 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. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.

製品仕様

You are viewing: N2016 Change Configuration

選択製品の構成品内容

Item Part # Size

FRB-LgBiT Control Vector

N200A 1 × 20μg

FKBP-SmBiT Control Vector

N201A 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

See Protocol for detailed storage recommendations.

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.

No transfer of this product is allowed. Researchers may use this product in their own research and they may transfer derivatives to others for research use provided that at the time of transfer a copy of this label license is given to recipients and recipients agree to be bound by the terms of this label license.

Researchers shall have no right to modify or otherwise create variations of the nucleotide sequences of the subunits of NanoBiT® Technology, except that researchers may create fused gene sequences at the termini of the subunits of NanoBiT® Technology. No other use of this product or its derivatives is authorized without the prior express written consent of Promega.

In addition, researchers must either:
(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 either:
(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 has no right to modify, derivatize, genetically engineer or otherwise create variations of the NanoBiT® Control Vectors, except that researcher may propagate and store NanoBiT® Control Vectors for use in NanoBiT® assays. 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. 9,797,889, 9,797,890 and 11,493,504; European Pat. No. 2970412; Japanese Pat. Nos. 7280842 and 7532562; and other patents pending.

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