Autophagy is a vital process in which cells degrade and recycle their own components via double-membrane vesicles. It plays a key role in maintaining cellular homeostasis, responding to stress, and regulating disease pathways. Accurate and sensitive autophagy detection helps uncover disease mechanisms, screen modulators, and accelerate drug development.

Service Offerings

1. Fluorescent LC3 Virus–Based Detection

• Single-Fluorescent LC3 System

LC3 is fused to a single fluorescent protein (e.g., GFP) to label autophagosomes. High-resolution fluorescence imaging quantifies autophagosome number and distribution.

• Dual-Fluorescent LC3 System

LC3 is fused to two spectrally distinct fluorescent proteins (e.g., GFP and RFP). This enables discrimination between autophagosome precursors and autolysosomes and allows dynamic measurement of autophagic flux.

2. Protein and Cell-Population Analysis

• Western Blot Analysis

Quantifies LC3-I to LC3-II conversion and levels of autophagy markers such as p62 to assess autophagy activity.

• Flow Cytometry

Uses fluorescent labels or specific antibodies to analyze whole-cell or subpopulation shifts, rapidly evaluating autophagy signals across cell populations.

3. Key Readouts

• Autophagosome Count: Number of autophagosomes per cell measured by imaging.
• LC3-II/LC3-I Ratio: Western blot quantitative ratio reflecting progression of autophagy.
• p62 Level: Changes in p62 protein indicate autophagic degradation efficiency.
• Fluorescence Intensity: Signal intensity from imaging or flow cytometry quantifies autophagic flux.

Key Advantages

Our platform integrates multiple detection modes—fluorescence imaging, Western blot, and flow cytometry—to cover every stage of autophagy from initiation to degradation. We ensure stable, low-background signals with high reproducibility. A multidisciplinary team supports your project end to end, from virus construction and cell modeling to comprehensive data analysis. We also offer flexible customization of vector design, cell lines, and treatment conditions to meet your specific needs.

Typical Applications

• High-throughput screening of small molecules or natural products targeting autophagy
• Evaluation of gene knockdown/knockout effects on autophagy
• Investigation of interactions and mechanisms of autophagy-related proteins
• Dynamic monitoring of autophagy in disease models (e.g., neurodegeneration, cancer)
• Lead optimization of autophagy modulators

Considerations

• Cell Line Selection: Autophagy baseline varies among cell types; choose the most relevant model.
• Fluorescence Overlap: In dual-fluorescent assays, minimize spectral overlap by optimizing filters and excitation sources.
• Replicates & Statistics: Perform at least three biological replicates and appropriate statistical analyses to ensure data reliability.
• Biosafety: All virus and cell work complies with Biosafety Level 2 regulations to protect personnel and the environment.

For more information or to discuss a customized detection solution, please contact our technical support team.