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DALGreen - Autophagy Detection

Item # Unit Size
D675-10
20 nmol

For Research Use Only Products

∼ Features ∼

No need to express protein! Just add the Working Solution for fluorescent imaging.
  1. Very Simple Procedure. Just Add the Working Solution
  2. Can be used for live autophagy monitoring
  3. Excitation at 488 nm



Content : 20 nmol x 1
Storage conditions : Store at -20oC and protect from light
Shipping conditions : Ambient Temperature


  • Product Description
  • Simple Procedure
  • Comparison to LC3
  • Comparison to MDC

  • Product Description

    DALGreen is used to detect autophagy in live cells. Autophagy is an intracellular degradation system, where dysfunctional proteins and organelles are degraded. In this process, aggregated dysfunctional proteins are surrounded by the double membrane to form an autophagosome. DALGreen, which is a small hydrophobic molecule, passes the plasma membrane of live cells and is incorporated in the autophagosome. After a lysosome fuses with the autophagosome, the environment in the autolysosome become acidic. DALGreen fluoresce stronger as acidity increases. The quality of this dye enables live cell imaging with a fluorescence microscopy and quantitative assay by flow cytometry.



    Simple Procedure

    First, cells are plated for the assay. After discarding the supernatant and washing the cells with culture medium, DALGreen working solution is added. While incubating for 30 minutes, DALGreen passes the plasma membrane of live cells and is incorporated in the forming autophagosome (Fig.1). After the 30 minutes incubation, supernatant is removed and the cells are washed. Medium containing autophagy-inducing agent is added to the cells. After incubating, observation can be made via fluorescence microscope or flow cytometer. For more detailed procedure, please refer to the manual.



    Comparison to LC3


    Method of autophagy induction
    Control: incubated in the cell media for 6 hours
    Starved: incubated in media without amino acid for 6 hours

    Condition of DALGreen Imaging
    Cell line: HeLa
    Wavelength: Ex. 488 nm/ Em. 500-563 nm
    Scale bar: 20 μm


    Comparison to MDC

    DALGreen can be used for live autophagy monitoring because Working Solution is added prior to inducing autophagy unlike MDC.




    References

    1) Iwashita, H., Sakurai, H. T., Nagahora, N., Ishiyama, M., Shioji, K., Sasamoto, K., Okuma, K., Shimizu, S. and Ueno, Y. (2018), Small fluorescent molecules for monitoring autophagic flux. FEBS Letters. doi: 10.1002/1873-3468.12979