ViaScript® LgBiT mRNA Delivery System
- HiBiTタグ付きタンパク質のライブセル動態解析が向上
- 細胞集団全体で効率的かつ均一な発現が得られる
- DNAプラスミドよりも速やかに発現するため、より早く結果を取得
Catalog Number:
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Size
Catalog Number: NE1120
Catalog Number: NE1130
Catalog Number: NE1140
内在性タンパク質発現をリアルタイムで定量化
HiBiTは11アミノ酸からなるエピトープタグで、LgBiTと自発的に結合し、明るい発光を示すNanoBiT®ルシフェラーゼを形成します。CRISPR/Cas9によるゲノム編集を用いてHiBiTを標的遺伝子座に導入することで、内在性レベルでのタンパク質の解析が可能になります。 ViaScript® LgBiT mRNA Delivery SystemとNano-Glo® Extended Live Cell Substratesを組み合わせることで、HiBiTタグ付きタンパク質をライブセルのまま、単一ウェルで動態的に定量することができます。
HiBiT を用いた動態解析がもたらす利点
- 単一時点測定では検出が困難な効果の観察（例：フック効果）
- タンパク質の分解・回復速度など、化合物の重要な特性を正確に把握でき、創薬研究やスクリーニング、最適化に活用可能
- 同一ウェルを用いた繰り返し測定による高い再現性.
- 各時間点用のウェルを分ける必要がないことによる時間と試薬の節約
ViaScript® LgBiT mRNA Delivery Systemを用いることで、より迅速かつ均一なLgBiT発現が可能となり、ライブセルHiBiT動態アッセイにおいて、代表性と精度の高いデータが得られます。
均一な発現による一貫性と信頼性の高い結果の実現
ViaScript® LgBiT mRNA Delivery Systemは、LgBiT のより均一かつ効率的な導入を可能にし、生物学的により関連性の高い細胞株において、HiBiT タグ付き目的タンパク質を内在性レベルで解析することができる。
22Rv1 AR-HiBiT クローン細胞株に対し、ViaScript® LgBiT mRNA Delivery System（左）または LgBiT 発現ベクター（右）を用いて一晩トランスフェクションを行った。細胞には、コントロール（上）または 1µM の ARV-110 PROTAC（下）を6時間処理し、AR の分解を誘導した。Nano-Glo® Live Cell Substrate 添加後、GloMax® Galaxy にてイメージ取得を行った。
エンドポイントアッセイでは見逃される効果の可視化
ViaScript® LgBiT mRNA Delivery Systemは、HiBiT CRISPRノックイン細胞株におけるライブセルでのタンパク質量の定量性能を向上させ、エンドポイントアッセイを超えたモデルおよび化合物に関する深い知見を提供する。上図に示すように、HiBiT を用いた動態的な標的タンパク質分解アッセイでは、タンパク質分解速度の精密な測定、フック効果の同定（中央パネルの矢印）、およびタンパク質の回復過程の特徴付けが可能である。
より迅速に結果を取得
ViaScript® LgBiT mRNA Delivery Systemによるトランスフェクションでは、DNAプラスミドを用いた場合よりも早く HiBiT シグナルがピークに達するため、実験をより早く開始でき、データを迅速に取得することができます。
生物学的により関連性の高い細胞株でアッセイを実施
The ViaScript® LgBiT mRNA Delivery Systemは、さまざまな細胞株において DNA トランスフェクションを上回る高い導入効率と低い細胞毒性を示す。HiBiT CRISPR ノックイン細胞株と mRNA トランスフェクションを組み合わせることで、より疾患の病態を反映した細胞環境で解析を行うことが可能となり、代表性の高い結果が得られ、研究の成功を後押しする。
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
eGFP mRNA |
EV495A | 1 × 100μl | |
ViaScript® Buffer |
EV100A | 1 × 11ml | |
ViaScript® Reagent |
EV496A | 1 × 0.1ml | 1mg/ml |
LgBiT mRNA |
N421A | 1 × 100μl |
NanoLuc® and NanoBiT® Reporter mRNAs Limited Use Label License
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 by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; and (2) resale of the product, whether or not such product is resold for use in research. Notwithstanding the foregoing, researcher may use this product in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the product. Researcher has no right to reverse engineer this product.
In addition, researcher 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, researcher 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.
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. 9,797,889 and other patents pending.
Patent Pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
ViaScript® Buffer |
EV100A | 1 × 11ml | |
ViaScript® Reagent |
EV496A | 1 × 0.1ml | 1mg/ml |
LgBiT mRNA |
N421A | 1 × 100μl |
NanoLuc® and NanoBiT® Reporter mRNAs Limited Use Label License
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 by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; and (2) resale of the product, whether or not such product is resold for use in research. Notwithstanding the foregoing, researcher may use this product in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the product. Researcher has no right to reverse engineer this product.
In addition, researcher 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, researcher 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.
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. 9,797,889 and other patents pending.
Patent Pending.
選択製品の構成品内容
| Item | Part # | Size | Concentration |
|---|---|---|---|
ViaScript® Buffer |
EV100A | 5 × 11ml | |
ViaScript® Reagent |
EV496A | 5 × 0.1ml | 1mg/ml |
LgBiT mRNA |
N421A | 5 × 100μl |
NanoLuc® and NanoBiT® Reporter mRNAs Limited Use Label License
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 by a party in exchange for consideration, including, but not limited to (1) use in further product manufacture; and (2) resale of the product, whether or not such product is resold for use in research. Notwithstanding the foregoing, researcher may use this product in provision of services, information or data to third parties in exchange for consideration, provided that researcher does not transfer the product. Researcher has no right to reverse engineer this product.
In addition, researcher 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, researcher 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.
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. 9,797,889 and other patents pending.
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GloMax® Discover System
発光・蛍光・吸光度測定に対応した高性能マイクロプレートリーダー
GM3000
