Lyso-Tracker Green, 50 μL

Product Information

Product Description/Introduction

Lyso-Tracker Green (DND-26), a lysosomal green fluorescent probe, can be used to specifically stain lysosomes in living cells. The probe is composed of a fluorescent group and connected weak bases, which can freely penetrate into cells and aggregate towards lysosomes with a weakly acidic internal environment, thus realizing the labeling of lysosomes.

Storage and Shipping Conditions

Ship with dry ice; Store at -20 away from light for 12 months.

Product Content

Assay Protocol / Procedures

1. Assay working solution preparation:

1.1. This product is a 1 mM storage solution that is warmed back to room temperature prior to experimentation and centrifuged briefly at low speed to ensure that the reagents sink to the bottom of the tube;

1.2. It is diluted by buffer (Hanks, PBS, etc., Hanks is recommended) or basal medium (without serum), into a lysosomal assay working solution of 50-100 nM ( Lyso-Tracker Green Fluorescent Probe Working Concentration, can be adjusted to some extent depending on the situation, but in order to avoid too high a concentration of probe, which will lead to a decrease in the specificity of the probe, resulting in false positives and so on, it is recommended that a lower concentration be used);

2. Cell staining (the following protocol is for adherent cells; centrifugation is required for suspension cell exchange)

2.1. Normal or treated cells are washed 1-2 times with buffer for 3-5 min each time;

2.2. Add pre-warmed lysosomal assay working solution and incubate in a cell culture incubator at 37 for 30-60 min (it is recommended to observe the fluorescence effect under a fluorescence microscope first, if the effect is not obvious, the incubation time can be extended appropriately);

2.3. Remove the incubation solution and wash with buffer solution 2-3 times for 3-5 min each time;

3. Microscopic examination：Ready for observation under a fluorescence microscope, FITC filter was selected with excitation wavelength of 504 nm and emission wavelength of 511 nm..

Note

1. Due to the different types and states of the cells, the working concentration of the dye, and the incubation time, can be adjusted appropriately; The study of the internalization kinetics of the probes has shown that it takes only a few seconds for the probes to enter the living cells. It is important to note that the probe causes an "alkalinization effect" in the lysosome, and therefore prolonged incubation may cause an increase in lysosomal pH.

2. The dye needs to perform lysosomal localization in the state of living cells, and it is not suitable for subsequent cell fixation operations.

3. When staining and washing live cells, the assay working solution and washing buffer need to be pre-warmed at 37°C. Staining and washing are accomplished by incubation in an incubator so as not to affect the morphology of the cells by temperature difference stimulation.

4. The fluorescence of this probe is easily quenched, so it should be photographed as soon as possible after staining, and the whole staining process needs to be operated away from light.

5. Probe master mixes should be stored in separate containers to avoid repeated freezing and thawing.To avoid waste, the staining solution is prepared as needed, ready to use.

6. For your health and safety, please wear lab coat and gloves during operation.

For Research Use Only!

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