Nano-Glo® HiBiT Dual-Luciferase® Reporter System
Multiplexed Detection of HiBiT-Tagged Proteins and Firefly Luciferase in the Same Sample
- Rapidly identify specific and non-specific effects of compounds
- Compatible with batch processing and automation
Catalog Number:
Size
Catalog Number: NE2010
Catalog Number: NE2020
同一サンプルからのホタルルシフェラーゼとHiBiTタグ付きタンパク質の連続測定
Nano-Glo® HiBiT Dual-Luciferase® Reporter System（HiBiT NanoDLR™）は、1つのサンプルからホタルルシフェラーゼ（Fluc）とHiBiTタグ付きタンパク質の発光を連続測定できる「添加→測定→添加→測定」形式のアッセイシステムです。Flucは構成的に発現するコントロールとして使用され、HiBiTタグ付き標的タンパク質の特異的な変動ではなく、細胞生存率やタンパク質発現に対する化合物の非特異的な影響を評価するための指標となります。これにより、タンパク質調節スクリーニングや関連研究におけるデータの解釈をサポートします。
11アミノ酸からなるHiBiTペプチドタグは、CRISPR/Cas9ゲノム編集とFlucの異所性発現を組み合わせることで、目的のタンパク質に付加することができます。あるいは、バイシストロニック型HiBiT導入ベクターを使用することで、HiBiT融合タンパク質と構成的に発現するFlucの両方を同一mRNA上で発現させることも可能です。
Simplify Your Workflow
Explore premade solutions to simplify your workflow—whether you need ready-to-use firefly expression vectors, HiBiT-tagged protein vectors or prebuilt knock-in CRISPR cell lines designed for reliable expression and detection.
HiBiT NanoDLR™ のワークフロー
ホタルルシフェラーゼ（Fluc）は、LgBiTを含むONE-Glo™ EX Reagentを添加することで最初に検出される（1）。その後、NanoDLR™ Stop & Glo® Reagentを細胞に添加することで、Flucシグナルを消光すると同時に、NanoBiT®（HiBiT + LgBiT）シグナルが検出される（2）。
Flucシグナルでタンパク質レベルへの特異的／非特異的影響を識別
2種のPROTACによるHiBiT-BRD4の選択的分解と一般的な細胞毒性の比較 HEK293細胞に、弱いTKプロモーター下でHiBiT-BRD4-IRES-luc2カセットを発現するコンストラクトを一過性導入した。細胞には、MZ1（パネルA）、dBET1（パネルB）、またはstaurosporine（パネルC）を段階希釈して処理し、その後、溶解型HiBiT NanoDLR™ Assayを用いてFluc（青）およびHiBiT（赤）シグナルを測定した。MZ1およびdBET1は、BRD4をそれぞれ異なるE3リガーゼ複合体に選択的に誘導して分解させるのに対し、staurosporineは一般的な細胞毒性を引き起こし、HiBiT-BRD4およびホタルルシフェラーゼ（Fluc）の両方の減少を招いた。
7桁のダイナミックレンジで定量可能
発現レベルにかかわらずタグ付きタンパク質を正確に定量可能な広範な直線ダイナミックレンジ HiBiT NanoDLR™ Assay は、内在性レベルで発現する低発現タンパク質であっても定量可能である。
Protocols
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
ONE-Glo™ EX Luciferase Assay Buffer |
E632A | 1 × 10ml |
ONE-Glo™ EX Luciferase Assay Substrate |
E633A | 1 × 1 vial |
NanoDLR™ Stop & Glo® Buffer |
N142A | 1 × 10ml |
NanoDLR™ Stop & Glo® Substrate |
N143A | 1 × 100μl |
LgBiT Protein |
N401A | 1 × 0.1ml |
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. 9,487,814, European Pat. No. 2959010 and other patents and patents pending.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. No. 9,797,889 and other patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
ONE-Glo™ EX Luciferase Assay Buffer |
E632B | 1 × 100ml |
ONE-Glo™ EX Luciferase Assay Substrate |
E633B | 1 × 1 vial |
NanoDLR™ Stop & Glo® Buffer |
N142B | 1 × 100ml |
NanoDLR™ Stop & Glo® Substrate |
N143B | 1 × 1ml |
LgBiT Protein |
N401B | 1 × 1ml |
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. 9,487,814, European Pat. No. 2959010 and other patents and patents pending.
U.S. Pat. No. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
U.S. Pat. No. 9,797,889 and other patents pending.
Related Products
類似製品
Nano-Glo® HiBiT Blotting System
抗体を用いずにブロッティングメンブレン上の HiBiT タンパク質を迅速に検出する発光法
N2410
Nano-Glo® HiBiT Lytic Detection System
細胞内の HiBiT-タグ融合タンパク質総量を発光法により検出
N3030, N3040, N3050
Nano-Glo® HiBiT Extracellular Detection System
細胞表面や培地中に分泌されたHiBiT タグタンパク質を生細胞のまま発光検出
N2420, N2421, N2422
Anti-HiBiT Monoclonal Antibody
新たにHiBiTタグの抗体検出を可能にした特異的で高感度なモノクローナル抗体
N7200, N7210
