|Product Overview:||Lysosomes are membrane-bound organelles important for various cellular processes. They contain hydrolytic enzymes that are utilized in the metabolism of some biomolecules. The extracellular cargo (e.g. nutrients toxins) binds to the cell membrane and is subsequently transported into membrane-bound endosomes for further degradation by lysosomes while intracellular components are transported to lysosomes through autophagy. Lysosomal dysfunction is associated with many human conditions such as aging and neurodegenerative disease. Although the intracellular activity of lysosome is difficult to measure in living cells, We have developed a proprietary Lysosome-Specific Self-Quenched Substrate which has low background fluorescence, high signal to background ratio and is pH insensitive. The substrate, acting as endocytic cargo, can be taken up by cells and degraded within an endo-lysosomal vesicle. The fluorescent signal is recovered from the Self-Quenched Substrate. The fluorescence signal, generated by degradation, is proportional to the intracellular lysosomal activity and can be measured using a fluorescence microscopy and/or flow cytometry. Lysosomal Intracellular Activity Assay Kit (Cell-Based) includes Cytochalasin D, a cell-permeable inhibitor of endocytosis that serves as an experimental control. This easy-to-use, non-radioactive kit allows imaging and accurate measurement of de-quenching substrate in cultured cells.|
|Applications:||Measurement of lysosomal intracellular activity.
Elucidation of the mechanisms of endocytic pathway in living cells.
Screening compounds with anti-lysosomal intracellular activity.
|Kit Components:||Polyamine Assay Buffer; Polyamine Probe (in DMSO); Polyamine Enzyme Mix; Polyamine Developer; Sample Clean-Up Mix; Polyamine Standard|
|Detection method:||Flow Cytometry (FL1)/ Microscopy (Ex: 488 nm)|
|Features & Benefits:||Measurement of lysosomal intracellular activity.|