Autophagy/Cytotoxicity Dual Staining Kit

Autophagy is a critical cellular process that involves the degradation and digestion of intracellular components by the lysosome. This process not only enables cells to efficiently mobilize and recycle cellular constituents, but also prevents the accumulation of damaged organelles, misfolded proteins, and invading microorganisms.¹ Autophagy is a multi-step process that begins with the sequestration of cytoplasm, organelles, and proteins. These cellular components are sequestered by a double membrane, forming an autophagosome. The autophagosome then fuses with a lysosome to form an autolysosome, where the cellular material is then degraded.^2,3 Normal autophagy is essential for survival, differentiation, development, and homeostasis. Dysregulation of autophagy has been implicated in cancer, infection, aging, and degenerative diseases.² While autophagy most often acts to promote cell survival in response to stress, it can also promote cell death. The relationship between autophagy and apoptosis is complex. The two pathways share common stimuli, components, and can regulate the activity of each other.⁴ However, the specific factors and mechanisms that dictate the choice between autophagy and apoptosis remain unclear. Cayman’s Autophagy/Cytotoxicity Dual Staining Kit provides a convenient tool for studying the regulation of autophagy and cytotoxicity at the cellular level. The kit employs monodansylcadaverine (MDC), an autofluorescent substance incorporated into multilamellar bodies by both an ion trapping mechanism and the interaction with membrane lipids,⁵ as a probe for detection of autophagic vacuoles in cultured cells. Propidium iodide is used as a marker of cell death. Tamoxifen, a known inducer of autophagy, is included as a positive control.

Needed but not supplied: Please download the kit booklet to verify if UltraPure Water (Milli-Q or equivalent) or any other components are needed for this assay.

WARNING This product is not for human or veterinary use.

1. Jin, S., and White, E. Tumor suppression by autophagy through the management of metabolic stress. Autophagy 4(5), 563-566 (2008).

2. Levine, B., and Kroemer, G. Autophagy in the pathogenesis of disease. Cell 132, 27-42 (2008).

3. Munafó, D.B., and Colombo, M.I. A novel assay to study autophagy: Regulation of autophagosome vacuole size by amino acid deprivation. J. Cell Sci. 114, 3619-3629 (2001).

4. Mizushima, N., Levine, B., Cuervo, A.M., et al. Autophagy fights disease through cellular self-digestion. Nature 451, 1069-1075 (2008).

5. Niemann, A., Takatsuki, A., and Elsässer, H.P. The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probe. J. Histochem. Cytochem. 48(2), 251-258 (2000).