Autophagy LC3 HiBiT Reporter Assay System
Multimodal Assay for Detection of Autophagic Flux
- Quantitative, unambiguous, scalable LC3 reporter assay
- No-wash, add-mix-measure protocol
- Blotting and imaging formats for confirmatory readings
Catalog Number:
Choose a reporter cell line or reporter vector
Catalog Number: GA1040
Catalog Number: GA1050
Catalog Number: GA2550
Simple Detection of Autophagic Flux and Autophagosome Formation
シンプルな生物発光によるオートファジーフラックス検出
Autophagy LC3 HiBiT Reporter Assay System はオートファジーフラックスを定量的に測定する発光 LC3 レポーターを用いたプレートベースのアッセイです。簡便な1ステッププロトコールで洗浄操作や複雑なイメージングまたはフローサイトメトリーシステムは不要です。構築済みの2種類のレポーター細胞株 (HEK293 または U2OS) またはご自身の細胞株で安定トランスフェクションするためのレポーターベクターからお選びいただけます。
How to Measure Autophagy Using the LC3 Assay
測定原理
Autophagy LC3 HiBiT Reporter はヒト LC3タンパク質に HiBiTタグを融合させたものです。オートファジーが誘導されると LC3 レポータータンパク質がオートファゴゾーム中に取り込まれ、分解されます。LgBiT タンパク質と基質を含む Nano-Glo® HiBiT Lytic Reagent を添加すると LgBiT が HiBiTタグと相互作用し、 NanoBiT® 酵素が再構成されて光を生じます。この発光シグナルは HiBiT-tagged LC3 レポーターのレベルと7桁以上の直線性で相関します。
1 Assay for 3 Different Methods: Plate Reader, Protein Blotting and Imaging
Plate Reader Data: Quantify Autophagy Inducer/Inhibitor Effect
データ例
このオートファジーレポーターアッセイから得られる値は定量性があり解釈も容易です。オートファジーフラックスの上昇はオートファジーレポーター分解を促進し、発光シグナルを減少させます。反対に、オートファジーの阻害はレポーターレベルおよび発光シグナルを増加させます。
オートファジーの誘導：シグナルは減少する
オートファジーの阻害：シグナルは増加する
Protein Blotting Data: Measure LC3-I to LC3-II Conversion
Imaging Data: Visualize LC3 Localization to the Autophagosome
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
Nano-Glo® HiBiT Lytic Detection System |
N3030 | 1 × 10ml | View Product |
HEK293 Autophagy LC3 HiBiT Reporter Cell Line |
GA104A | 1 × 1 each |
NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If the 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.
This product may not be further sold or transferred by the researcher and may be used only by the researcher, and then only for research use, which includes, but is not limited to drug discovery and development. No other commercial use is allowed. "Commercial use" means any and all uses of this product by researcher for monetary or other consideration, including, but not limited to, (1) use in further product manufacture; (2) use in provision of services, information or data to unaffiliated third parties, and (3) resale of this product for any use. Researcher has no right to modify, derivatize, genetically engineer or otherwise create variations of the nucleotide sequence encoding the HiBiT and HaloTag® peptides which sequences have been stably transfected within the cells. Researcher may genetically engineer the cell line using exogenous nucleic acid sequences of the researcher’s choosing and researcher may propagate and store the cells for long-term use. In addition, researcher must use Nano-Glo® HiBiT Lytic Detection System purchased from Promega for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega's. For uses of HaloTag® Technology in this product, researchers must either: (1) use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties, or (2) contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
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 REGARDS TO THIS 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. Nos. 9,797,889, 9,797,890 and 11,493,504; European Pat. Nos. 2970412 and 3783011; Japanese Pat. Nos. 7280842 and 7532562; and other patents pending.
HEK293 cells were obtained under license from AdVec Inc.
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
U2OS Autophagy LC3 HiBiT Reporter Cell Line |
GA105A | 1 × 1 each | |
Nano-Glo® HiBiT Lytic Detection System |
N3030 | 1 × 10ml | View Product |
NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. BY USE OF THIS PRODUCT, RESEARCHER AGREES TO BE BOUND BY THE TERMS OF THIS LIMITED USE LABEL LICENSE. If the 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.
This product may not be further sold or transferred by the researcher and may be used only by the researcher, and then only for research use, which includes, but is not limited to drug discovery and development. No other commercial use is allowed. "Commercial use" means any and all uses of this product by researcher for monetary or other consideration, including, but not limited to, (1) use in further product manufacture; (2) use in provision of services, information or data to unaffiliated third parties, and (3) resale of this product for any use. Researcher has no right to modify, derivatize, genetically engineer or otherwise create variations of the nucleotide sequence encoding the HiBiT and HaloTag® peptides which sequences have been stably transfected within the cells. Researcher may genetically engineer the cell line using exogenous nucleic acid sequences of the researcher’s choosing and researcher may propagate and store the cells for long-term use. In addition, researcher must use Nano-Glo® HiBiT Lytic Detection System purchased from Promega for all luminescence assays using this product or contact Promega to obtain a license for use of this product with reagents other than Promega's. For uses of HaloTag® Technology in this product, researchers must either: (1) use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties, or (2) contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
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 REGARDS TO THIS 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. Nos. 9,797,889, 9,797,890 and 11,493,504; European Pat. Nos. 2970412 and 3783011; Japanese Pat. Nos. 7280842 and 7532562; and other patents pending.
選択製品の構成品内容
| Item | Part # | Size | Available Separately |
|---|---|---|---|
Autophagy LC3 HiBiT Reporter Vector |
GA255A | 1 × 20μg | |
Nano-Glo® HiBiT Lytic Detection System |
N3030 | 1 × 10ml | View Product |
NOT FOR MEDICAL DIAGNOSTIC USE. FOR IN VITRO USE ONLY. 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, which includes, but is not limited to drug discovery and development, 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. No other commercial use is allowed. "Commercial use" means any and all uses of this product by researcher for monetary or other consideration, including but not limited to, (1) use in further product manufacture; (2) use in provision of services, information or data to unaffiliated third parties, and (3) resale of this product for any use.
Researchers shall have no right to modify or otherwise create variations of the nucleotide sequences that encode the HiBiT and HaloTag® peptides. 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® HiBiT Lytic Detection System-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.
For uses of HaloTag® Technology in this product, researchers must either: (1) use Promega HaloTag® ligands, which can be modified or linked to Promega or customer-supplied moieties, or (2) contact Promega to obtain a license if Promega HaloTag® ligands are not to be used.
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. Nos. 9,797,890 and 10,648,971; Japanese Pat. Nos. 7270842 and 7532562; and other patents and patents pending.
U.S. Pat. Nos. 8,420,367, 8,748,148, 9,593,316 and 9,873,866; European Pat. Nos. 2374875, 2492342 and 2502990; and Japanese Pat. Nos. 5680302 and 5840117.
U.S. Pat. Nos. 9,797,889, 9,797,890 and 11,493,504; European Pat. Nos. 2970412 and 3783011; Japanese Pat. Nos. 7280842 and 7532562; and other patents pending.
Resources
技術記事
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