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Cell Meter™ Autophagy Assay Kit
Blue Fluorescence
Autophagy is an evolutionarily conserved degradation process that targets long-lived proteins, organelles, and other cytoplasmic components for degradation via the lysosomal pathway. The autophagy pathway is complementary to the action of the ubiquitin-proteasome pathway which typically degrades short-lived proteins. Activation of the autophagy pathway is required for multiple cellular roles, including survival during starvation, the clearance of intracellular components, development, and immunity. In the absence of stress, autophagy serves a house-keeping function, removing damaged organelles and cellular components preventing cytotoxic effects. Decreases and defects in autophagy have been implicated in multiple diseases, for example Huntingtons, Alzheimers, and Parkinsons. In terms of cancer development, autophagy seems to play multiple roles. Decreased or absent expression of certain autophagy proteins, such as Beclin-1 and Bif-1, increases tumor susceptibility in mice while the overexpression of these proteins can repress cancer cell growth. However, autophagy is critical for the survival of cancer cells within the nutrient poor and hypoxic core of solid tumors. Cell Meter™ Autophagy Kit employs Autophagy Blue™ as a specific autophagosome marker to analyze the activity of autophagy. The assay is optimized for direct detection of autophagy in both detached and attached cells. The kit provides all the essential components for the assay protocol. Cell Meter™ Autophagy Kit is suitable for fluorescence microscope, fluorescence microplate reader.
Autophagy Blue&trade; labeled vesicles are induced by starvation in HeLa cells using Cell Meter&trade; Autophagy Assay Kit. HeLa cells were incubated in a regular DMEM medium as control (A) or in a serum-depleted medium as autophagy treatment (B) for 16 hours. Both control cells and starved cells were incubated with Autophagy Blue&trade; working solution for 30 minutes in a 37 &deg;C, 5%&nbsp; CO<sub>2&nbsp;</sub>incubator, and then washed four times with wash buffer. Cells were imaged immediately under a fluorescence microscope with a DAPI channel. Autophagy is indicated by bright blue dot staining of autophagic vacuoles (B).
Autophagy Blue&trade; labeled vesicles are induced by starvation in HeLa cells using Cell Meter&trade; Autophagy Assay Kit. HeLa cells were incubated in a regular DMEM medium as control (A) or in a serum-depleted medium as autophagy treatment (B) for 16 hours. Both control cells and starved cells were incubated with Autophagy Blue&trade; working solution for 30 minutes in a 37 &deg;C, 5%&nbsp; CO<sub>2&nbsp;</sub>incubator, and then washed four times with wash buffer. Cells were imaged immediately under a fluorescence microscope with a DAPI channel. Autophagy is indicated by bright blue dot staining of autophagic vacuoles (B).
protocol
Protocol
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CatalogSize
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Quantity
23000200 Tests
Price
 
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
Instrument settings
Fluorescence microscope
ExcitationDAPI channel
EmissionDAPI channel
Recommended plateBlack wall/clear bottom
Fluorescence microplate reader
Excitation330 nm
Emission520 nm
Cutoff475 nm
Recommended plateBlack wall/clear bottom
Instrument specification(s)Bottom read mode
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Page updated on October 12, 2025