Item number Product concentration Ex/Em save

IF1830 Lysosomal dark red fluorescent probe 1 mM 649/665 -20 ℃,2 years

IF1840 Lysosomal red fluorescent probe 1 mM 577/590 -20 ℃,2 years

IF1850 Lysosome green fluorescent probe 1 mM 504/511 -20 ℃,2 years

Usage (for reference only)

1. Preparation of lysosomal probe working solution

(1) A small amount of lysosomal probe reserve solution (1mM) was added into the cell culture solution at the ratio of 1:10,000-1:20,000, so that the final concentration was 50-100 nM, and the lysosomal probe working solution was formed after mixing.

(2) The lysosomal probe working solution can be pre-warmed at 37℃ before use.

2. Fluorescent labeling of lysosomes

(1) The cell medium was removed, and after 1×PBS was cleaned once, the lysosomal probe working solution prepared in Step 1 was added, and the cells were incubated at 37℃ for 30 min and 2 h. The incubation time of different cells was different, and it was recommended to adjust according to the staining effect.

(2) The lysosomal probe working solution was removed, and after three times of cleaning with 1×PBS, the fluorescence microscope was photographed for observation. If it is necessary to add Hoechst 33342 to re-stain, it is recommended to use Hoechst 33342 with a concentration of 10 μg/mL and incubate at 37℃ for 5 min to remove the dye. Take pictures after 1×PBS cleaning.

Matters needing attention

1. If the staining effect is not good, the concentration of the lysosomal probe can be increased, or the dyeing time can be extended appropriately within the recommended time range.

2. In order to reduce the dyeing background, use a lower concentration of dye as much as possible.

3. Take pictures quickly, because the dye is easy to quench with the increase of the photo time.