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
Rilevazione semplice del flusso autofagico e della formazione di autofagosomi
Il sistema Autophagy LC3 HiBiT Reporter Assay utilizza un reporter LC3 luminescente per misurare quantitativamente il flusso autofagico. È possibile scegliere tra una delle due linee cellulari reporter preparate (HEK293 o U2OS) o trasfettare stabilmente il vettore reporter nella linea cellulare di propria scelta. Con un unicosaggio è possibile verificare i risultati utilizzando tre metodi diversi: lettore di piastre, protein blotting e imaging.
Come misurare l'autofagia con il saggio LC3
L'Autophagy LC3 HiBiT Reporter è costituito da un tag HiBiT fuso con la proteina LC3 umana, collegato da uno spaziatore HaloTag®. Quando l'autofagia è indotta, le proteine reporter LC3 vengono catturate negli autofagosomi e degradate. Con l'aggiunta del reagente litico Nano-Glo® HiBiT contenente la proteina LgBiT e il substrato, LgBiT interagisce con il tag HiBiT per ricostituire l'enzima luminoso e luminescente NanoBiT®. Il segnale luminescente è direttamente proporzionale al livello del reporter LC3 marcato HiBiT per oltre sette ordini di grandezza. Ulteriori moduli di rilevamento sono il blotting HiBiT e l'imaging con ligandi HaloTag®.
1 test per 3 diversi metodi: Lettore di piastre, blotting di proteine e imaging
Dati del lettore di piastre: Quantificare l'effetto degli induttori/inibitori dell'autofagia
Con un semplice lettore di piastre, è possibile quantificare facilmente i livelli di autofagia rilevando la luminescenza. Un aumento del flusso autofagico accelererà la degradazione del reporter dell'autofagia, con conseguente diminuzione del segnale luminescente. Al contrario, l'inibizione dell'autofagia aumenterà i livelli del reporter e il segnale di luminescenza.
Induzione dell'autofagia: Segnale ridotto
Inibizione dell'autofagia: Aumento del segnale
Dati sul blotting delle proteine: Misurare la conversione da LC3-I a LC3-II
Dati di imaging: Visualizzazione della localizzazione di LC3 nell'autofagosoma
Specifications
Catalog Number:
What's in the box
| Item | Part # | Choose a 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 |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.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.
What's in the box
| Item | Part # | Choose a 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 |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.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.
What's in the box
| Item | Part # | Choose a size | Available Separately |
|---|---|---|---|
Autophagy LC3 HiBiT Reporter Vector |
GA255A | 1 × 20μg | |
Nano-Glo® HiBiT Lytic Detection System |
N3030 | 1 × 10ml | View Product |
SDS
Search for SDSCertificate of Analysis
Use Restrictions
For Research Use Only. Not for Use in Diagnostic Procedures.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.
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Frequently Used With
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Bioluminescent method for detecting the total amount of HiBiT-tagged proteins in the cell.
N3030, N3040, N3050
Nano-Glo® HiBiT Blotting System
Luminescent method for rapid detection of HiBiT-tagged proteins on blots—no antibodies needed.
N2410
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Dyes for fluorescent labeling, confocal imaging and high-resolution single-molecule imaging studies of HaloTag® fusion proteins in living cells.
HT1110, HT1100, HT1010, CS315102, HT1020, HT1030, HT1040, HT1050, HT1060, HT1070, HT1080, HT1090, HT1120, HT1130, CS3495C04, CS3495C05, CS3495C06, CS3495C07, GA1110, GA1111, GA1120, GA1121, G8581, G8582, G8272, G8273, G2801, G2802, G3221, G8251, G8252, G2991, G1001, G1002, G8471, G8472
FuGENE® HD Transfection Reagent
Enhanced performance in a range of cell types, including difficult-to-transfect cell lines.
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