Nano-Glo® HiBiT Extracellular Detection System
Quantify HiBiT-Tagged Proteins Expressed on the Cell Surface
- Specific, live-cell detection of extracellular expressed or secreted proteins
- Simple add-mix-read assay format—no antibodies required
- Quantitate over 7 logs of linear dynamic range
Catalog Number:
Size
Catalog Number: N2420
Catalog Number: N2421
Catalog Number: N2422
細胞表面のタンパク質発現をシンプルに発光検出
Nano-Glo® HiBiT Extracellular Detection System は細胞表面あるいは分泌された発現タンパク質をシンプルな "添加 - 混和 - 測定" アッセイフォーマットで数分以内に測定することができます。
細胞非破壊性の Nano-Glo® HiBiT Extracellular Detection Reagent を用いて 11アミノ酸で構成される HiBiT ペプチドを有し細胞外に発現するあらゆるタンパク質を特異的に定量することができます。検出試薬には HiBiT タグと親和性を持ち、明るい NanoBiT® 発光酵素として再構成される相補的なポリペプチド LgBiT を含みます。
発光は細胞外に存在する HiBiT タンパク質量に比例して7桁以上の直線性を有しています。
【日本語製品マニュアル・簡易マニュアルページは こちら】
※日本語マニュアルは英語版を翻訳したものですが、常に更新されるものではありません。最新のマニュアルについては英語版（以下の “プロトコール” を参照）をご覧ください。
【Q&A】 HiBiT
簡便な "添加 - 混和 - 測定" アッセイワークフロー
Nano-Glo® HiBiT Extracellular Detection Reagent を加え、混和して発光を測定するだけで細胞外に発現した HiBiT タグ タンパク質量を直接的に測定することができます。
Nano-Glo® HiBiT Detection Reagent には酵素基質と細胞膜非透過性の LgBiT ペプチドが含まれます。細胞表面上あるいは培地中に分泌された 発現 HiBiT タグタンパク質だけが細胞外の LgBiTと相補し NanoBiT® 発光酵素として再構成されます。
定量性の高い Nano-Glo® HiBiT Extracellular Detection System は標準的な抗体ベースの検出法に比べて操作ステップ数が飛躍的に少なくなるため時間を節約でき、精度も改善されます。
定量アッセイの開発
- 受容体のインターナリゼーション
- 受容体のリサイクリング
- タンパク質またはサイトカインの分泌
- 表面タンパク質のトラフィッキング
数分以内に受容体インターナリゼーションを測定
Nano-Glo® HiBiT Extracellular Detection System は受容体のインターナリゼーションのためのシンプルな定量アッセイ開発を可能にし、抗体ベースの方法よりも時間を節約でき、バラつきも低減することができます。最適化された検出試薬を用いることでタンパク質受容体との迅速な平衡化が可能となりウェル間のバラつきを最小限に抑え、生物学的変化を直ぐにキャッチすることができます。例えば、右図では4種類の完全または部分的アゴニストに暴露した際の β2-adrenergic receptor (ADRB2) のインターナリゼーション測定を行いました。
| Agonist | EC50 | Percentage Receptor Remaining on Surface |
|---|---|---|
| Isoproterenol | 50.9nM | 16% |
| Salbutamol | 161nM | 45% |
| Salmeterol | 1.04nM | 63% |
| Formoterol | 2.92nM | 16% |
実験スケールからHTSへ
マルチウェルプレートあるいは多検体処理への対応性によりNano-Glo® HiBiT Extracellular Detection System はより優れた柔軟性と利便性を提供します。そのため標準的な免疫検出法では不可能であった膜受容体や分泌タンパク質に関するスクリーニングアッセイの開発も可能です。
Internalization of HiBiT-ADRB2 in stably transfected HEK293 cells in 384-well plates
内在レベルのタンパク質を測定できる感度
11アミノ酸という HiBiT タグの小ささにより、クローニングを行わずに簡便にゲノム編集することもできます。 CRISPR/Cas9 を用いることで HiBiT タグを内在ローカスに厳密に挿入することができます。そのため、内在的に制御される膜受容体の機構を調べる上で、過剰発現によるアーティファクトを低減し、相互作用するタンパク質との化学量論的適性が維持されます。
Learn how to accurately measure protein surface expression with the Nano-Glo® HiBiT Extracellular and Lytic Assays in this whitepaper.
Protocols
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Extracellular Buffer |
N243A | 1 × 10ml |
Nano-Glo® HiBiT Extracellular Substrate |
N300A | 1 × 0.2ml |
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. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Extracellular Buffer |
N243B | 1 × 100ml |
Nano-Glo® HiBiT Extracellular Substrate |
N300B | 2 × 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. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Extracellular Buffer |
N243B | 10 × 100ml |
Nano-Glo® HiBiT Extracellular Substrate |
N300C | 5 × 4ml |
LgBiT Protein |
N401B | 10 × 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. 8,809,529, European Pat. No. 2635582, Japanese Pat. No. 5889910 and other patents and patents pending.
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Simplify Your Workflow
Explore premade solutions to simplify your workflow—whether you need ready-to-use HiBiT-tagged protein vectors or prebuilt knock-in CRISPR cell lines designed for reliable expression and detection.
