LysoTracker Red

• CAT Number: I044993
• CAS Number: 231946-72-8
• Molecular Formula: C20H24BF2N5O
• Molecular Weight: 399.25
• Purity: ≥95%

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].
Preparation of LysoTracker Red working solution
1. Working fluid configuration:
1.1 Restore the solution to room temperature and concentrate it at the bottom of the tube by instantaneous centrifugation.
1.2 Dilute 1 mM 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℃ 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℃ 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℃ 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.

Reference

[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.
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[2]. Pereira LC, et al. An autophagic process is activated in HepG2 cells to mediate BDE-100-induced toxicity. Toxicology. 2017;376:59-65.
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[3]. Nagahama M, et al. Cellular vacuolation induced by Clostridium perfringens epsilon-toxin. FEBS J. 2011;278(18):3395-3407.
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