LysoTracker Blue DND-22

LysoTracker blue DND-22 is a blue fluorescent dye (Ex/Em: 373/422 nm). LysoTracker blue DND-22 stains acidic regions in living cells. LysoTracker blue DND-22 is used in the researches of neurodegenerative diseases and leukemia^[1][2]. In Vitro:Guidelines (The following is our recommended protocol, which is provided as a guide only and should be modified to suit your specific needs).
Staining steps^[2]
1) Under the conditions of 37°C and 5% CO₂, U937 cells are cultured in IMDM medium containing 10% fetal bovine serum (HY-T1000), 100 units/mL Penicillin, and 50 μg/mL Streptomycin (HY-B1906).
2) Cells are treated with labeled oxLDL (18 μg/mL) or oxLDL-IC (24 μg/mL).
3) 50 nM LysoTracker Blue DND-22 is added to the cells 30 min before the end of the incubation with labeled oxLDL and oxLDL-IC.
4) After the incubation, cells are washed three times with serum-free IMDM to remove unbound dye.
5) Cells are suspended in 100 μL of IMDM, and then 10 μL aliquots are loaded into glass capillaries and sealed.
6) Cells are visualized using a confocal microscope with an excitation wavelength of 373 nm and an emission wavelength of 422 nm to observe lysosomal localization.