Nano-Glo® HiBiT Lytic Detection System
Accurately Quantify Any HiBiT-Tagged Protein Expressed in Cells
- Sensitive bioluminescent protein detection
- Simple add-and-read assay—no antibodies required
- Quantitate over 7 logs of linear dynamic range
Catalog Number:
Size
Catalog Number: N3030
Catalog Number: N3040
Catalog Number: N3050
シンプル、高感度な発光法によるタンパク質定量
Nano-Glo® HiBiT Lytic Detection System はシンプルな "添加 - 混和- 測定” だけのフォーマットを用いて高感度、広いダイナミックレンジで数分以内に細胞内タンパク質を定量することができます。
11 アミノ酸で構成される HiBiT ペプチドタグは簡単に目的タンパク質に付加することができ、細胞内の HiBiT タグタンパク質量をNano-Glo® HiBiT Lytic Detection Reagent を添加して測定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ともに NanoBiT® 発光酵素として再構成される ポリペプチド LgBiT が含まれています。発光強度は細胞ライセート中の HiBiT タグタンパク質量と7桁以上 直線的に比例します。長時間タイプの発光シグナルは数時間安定です。
【Q&A】 HiBiT
迅速で簡単なタンパク質定量 - 加えて、混ぜて、測定するだけ！
Nano-Glo® HiBiT Lytic Detection System を用いれば、細胞ライセート中の HiBiT タグタンパク質を簡便な "添加 - 混和 - 測定" アッセイで直接定量することができます。細胞を溶解すると同時に発光に必要な LgBiT タンパク質およびフリマジン基質を供給するNano-Glo® HiBiT Detection Reagent を HiBiTタグ タンパク質を発現する細胞に加えます。
LgBiT タンパク質と HiBiT タグとの高親和性の相互作用によりNanoBiT® 発光酵素が再構成されます。これにより標準的な抗体を用いた検出法よりも少ない操作ステップでより定量的な測定結果が得られます。
アッセイ法の開発例：
- 制御されたタンパク質発現
- 標的タンパク質の分解
- ウイルスの感染
- 遺伝子発現によるタンパク質レベルと RNA -seq 実験の確認
- プルダウンあるいは一過性トランスフェクション後のタンパク質定量
正確なタンパク質定量
発現レベルに関わらずにタンパク質存在量の変化を測定でき、タグタンパク質を広いダイナミックレンジで正確に定量できます。Nano-Glo® HiBiT Lytic Detection System により内在レベルの低いタンパク質量でも定量（検出限界 10-19 moles 以下）することできます。
Titrating purified HiBiT-tagged protein to demonstrate linear dynamic range
内在的に制御されたタンパク質を調べる
CRISPR/Cas9 ゲノム編集を用いれば HiBiT タグタンパク質を内在的に調節された条件下で発現させることができるため、過剰発現によるアーティファクトを低減し、内在性の結合パートナーや調節機構で適切な化学量論を維持することができます。
小さな HiBiTタグ（11アミノ酸）を目的タンパク質に付加する2つの方法：
- ご希望のプロモーターを選び、従来の組換え法を実施（べクターのトランスフェクション）
- シンプルでクローンが不要な CRISPR/Cas9 ゲノム編集プロトコールで内在ローカスに挿入（ゲノム編集）
ラボベンチから HTSへスケールアップ可能
シンプルなプロトコールと長い発光半減期 (3時間以上) により Nano-Glo® HiBiT Lytic Detection System はハイスループットアプリケーションにおける複数プレートのバッチ処理に理想的です。抗体ベースの方法では不可能なタンパク質調節に関するHTSアッセイを開発することができます。
Induced degradation of HiBiT-BRD4 in a CRISPR-derived pool of cells in 384-well plates
Real-World Example: Using CRISPR Gene Editing to Add HiBiT to Endogenous Genes
Learn how the tiny 11-amino-acid luminescent tag called HiBiT can quantify protein abundance even at endogenous levels. CRISPR-mediated gene editing was used add the HiBiT tag to several endogenous genes, including HIF1α, and quantified protein levels in response to various stimuli.
Protocols
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Lytic Substrate |
N246A | 1 × 0.2ml |
Nano-Glo® HiBiT Lytic Buffer |
N247A | 1 × 10ml |
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.
U.S. Pat. No. 9,797,889 and other patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Lytic Substrate |
N246B | 2 × 1ml |
Nano-Glo® HiBiT Lytic Buffer |
N247B | 1 × 100ml |
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.
U.S. Pat. No. 9,797,889 and other patents pending.
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
Nano-Glo® HiBiT Lytic Substrate |
N246C | 5 × 4ml |
Nano-Glo® HiBiT Lytic Buffer |
N247B | 10 × 100ml |
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.
U.S. Pat. No. 9,797,889 and other 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.
