Lentivirus-LC3 production service for Autophagy Flux Detection
Customized Lentivirus production
Introduction to AAV-LC3 production service for Autophagy Flux Detection
Premade LC3 Autophagy Biosensors Products and user manual
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Adeno associated virus(AAV) 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
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.
Genemedi has launched series of lentiviral packaging service of autophagy related biosensors, 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.
Besides, Genemedi has constructed several other fluorescent biosensors to study the different stages of autophagy flux, making the autophagy study much easier.
Properties
| Adenovirus Vector | |
|---|---|
| Quantity/Unit | Vials |
| Form | Frozen form |
| Suitable Types of Infection | In vivo infection in animals |
| Sipping and Storage Guidelines | Shipped by dry ice, stored at -80 ℃, effective for 1 year. Avoid repeatedly freezing and thawing. |
| Titer | > 1*10^8 TU/ml. |
Advantages
1. Pre-packaged, ready-to-use, fluorescently-tagged with monomeric GFP & RFP.
2. Higher efficiency transfection as compared to traditional chemical-based and other non-viral-based transfection methods. Ability to transfect dividing, non-dividing, and difficult-to-transfect cell types, such as primary cells or stem cells.
3. Non-disruptive towards cellular function.
4. Reveals changing cellular conditions in real time. Enables visualization under different cell/disease states in live cell and in vitro analysis.
Quality control description
Our optimized production of lentiviral vector and strict quality control systems supply customers with a high titer of functional recombinant lentiviral vectors. Two methods are employed to determine viral titers: physical titer (VP/mL) and functional titer (TU/mL). Physical titer is calculated by the level of protein, such as p24, or viral nucleic acid. The functional titer, a calculation of the active virus that can infect cells, is much less than the physical titer (100-1000 fold lower). The method we adopted is functional titer, which is an accurate solution for testing virus accurate activity and MOI. The physical titer can only reflect the number of virus particles, but not reflect the true viral activity, which will cause a large error in subsequent infection experiments.
Technical Documents
1. For further information about lentivirus administration and transduction, please see 
Lentivirus User Manual.
2. For further information about autophagy study, please see Lentivirus LC3 Autophagy Flux Detection User Manual.
Reference
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