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Promega Autophagy LC3 HiBiT Reporter Assay System

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For research use only and not for medical diagnostic use. The readout depends on a genetically expressed LC3 reporter and the tested cell context; controls and orthogonal imaging or blotting are needed to distinguish induction from blocked downstream turnover and exclude cytotoxic effects. Sources verified 2026-09-28: https://worldwide.promega.com/products/cell-health-assays/autophagy/autophagy-assay/ ; https://worldwide.promega.com/-/media/files/resources/protocols/technical-manuals/500/tm535-autophagy-lc3-hibit-reporter-assay.pdf?rev=d575b31d26fc4bcc92b006e835e76e27&sc_lang=en

Kind
Autophagic-flux research assay
Type of test
Luminescent LC3B-HiBiT reporter assay with optional imaging and blot confirmation
Sample
Live HEK293 or U2OS reporter cells, or a researcher-generated stable mammalian cell line expressing the reporter vector
Provider
Promega
How to get
Order HEK293 cells GA1040, U2OS cells GA1050, or reporter-vector system GA2550 from Promega; cell-culture facilities and a luminescence plate reader are required.
Details
The HaloTag-LC3B-HiBiT reporter enters autophagosomes and is degraded after autolysosome formation. Nano-Glo HiBiT reagent measures remaining intact reporter: increased flux lowers luminescence and decreased flux raises it. Reference inducer and lysosomal inhibitor controls support interpretation.