LysoTracker Red  (Synonyms: LysoTracker Red DND-99)

LysoTracker Red is a Red fluorescently labeled lysosomal probe with a maximum excitation/emission wavelength of 577/590 nm. The structure is composed of a fluorescein group and linked weak bases, which can freely cross the cell membrane and gather on spherical organelles. It is suitable for observing the internal biosynthesis and related pathogenesis of lysosomes^[1].

Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
Preparation of LysoTracker Red working solution
1. Working solution preparation:
1.1 Prepare LysoTracker Red stock solution with DMSO.
1.2 Dilute storage solution to the working solution using a medium or appropriate buffer (such as PBS) . The recommended working solution is 50-100 nM;
Note: Please adjust the concentration of the LysoTracker Red working solution according to the actual situation, and use it now.
2. Cell staining
2.1 For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
2.2 Add 1 mL of LysoTracker Red working solution, and then incubate at room temperature for 5-30 minutes.
2.3 Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.
2.4 Wash twice with PBS, 5 minutes each time.
2.5 Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

399.25

C₂₀H₂₄BF₂N₅O

231946-72-8

Solid

Brown to dark brown

590

577

O=C(CCC1=CC=C2C=C3C=CC(C4=CC=CN4)=[N]3[B+3]([F-])([N-]21)[F-])NCCN(C)C

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Price OT, et al. Quantitative fluorescence of 5-FU-treated fetal rat limbs using confocal laser scanning microscopy and Lysotracker Red. Cytometry A. 2003;53(1):9-21.  [Content Brief]

  [2]. Pereira LC, et al. An autophagic process is activated in HepG2 cells to mediate BDE-100-induced toxicity. Toxicology. 2017;376:59-65.  [Content Brief]

  [3]. Nagahama M, et al. Cellular vacuolation induced by Clostridium perfringens epsilon-toxin. FEBS J. 2011;278(18):3395-3407.  [Content Brief]