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Introduction-LC3 Autophagy Biosensors

Autophagy is a highly regulated homeostatic degradative process where cells destroy their own components via the lysosomal machinery and recycle them. This process plays both protective and deleterious roles in many diseases, including Alzheimer’s disease, aging, cancer, infection and Crohn’s disease. Elucidating the correlation between autophagy and apoptotic cell death has become the focus of a great deal of research. Members of the LC3 family play a key role in the maturation of the autophagosome. Lysosomal turnover of the autophagosome marker LC3-II reflects starvation-induced autophagic activity, and detecting LC3 by immunoblotting or immunofluorescence has become a reliable method for monitoring autophagy and autophagy-related processes, including autophagic cell death.

Autophagy is a self-degradative process in cell that is important for balancing sources of energy at critical times in development and in response to nutrient stress. For autophagy study, Genemedi supply autophagy biosensor, in which GFP and/or RFP tags are fused at the C-termini of the autophagosome marker LC3, allowing to detect the intensity of autophagy flux in real-time with more accuracy, clarity and intuitiveness. These biosensors provide an enhanced dissection of the maturation of the autophagosome to the autolysosome, which capitalizes on the pH difference between the acidic autolysosome and the neutral autophagosome. The acid-sensitive GFP will be degraded in autolysosome whereas the acid-insensitive RFP will not. Therefore, the change from autophagosome to autolysosome can be visualized by imaging the specific loss of the GFP fluorescence, leaving only red fluorescence.

For autophagy study, Genemedi supply autophagy flux biosensors encoded by AAV, Adenovirus and lentivirus, which can be used to observe autophagy flux and monitor the intensity of autophagy flux in real-time in vivo or in vitro. Welcome to email [email protected] for consultation.



Premade LC3 Autophagy Biosensors Products and user manual

Adeno associated virus

AAV-GFP-LC3 Autophagy Biosensor
AAV-mRFP-GFP-LC3 Autophagy Biosensor
 

Adenovirus

Adv-GFP-LC3 Autophagy Biosensor
Adv-mRFP-GFP-LC3 Autophagy Biosensor
 
 

Lentivirus

Lv-GFP-LC3 Autophagy Biosensor
Lv-mRFP-GFP-LC3 Autophagy Biosensor
 

LC3 Autophagy Biosensors User Manual

AAV-LC3 User Manual    pdf download
Adenovirus-LC3 User Manual    pdf download
Lentivirus-LC3 User Manual    pdf download


Confocal microscopic analysis of autophagosomes after amino acids deprivation

Taking advantage of RFP-GFP-LC3 and GFP-LC3 labeling system, in which GFP and/or RFP tags are fused at the C-termini of the autophagosome marker LC3, allowing to detect the intensity of autophagy flux in real-time with more accuracy, clarity and intuitiveness. These biosensors provide an enhanced dissection of the maturation of the autophagosome to the autolysosome, which capitalizes on the pH difference between the acidic autolysosome and the neutral autophagosome. The acid-sensitive GFP will be degraded in autolysosome whereas the acid-insensitive RFP will not.

Therefore, the change from autophagosome to autolysosome can be visualized by imaging the specific loss of the GFP fluorescence, leaving only red fluorescence.

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