LysoTracker Blue DND-22 (Synonyms: LysoTracker Blue DND-22; LysoTracker Red DND-99; Lysotracker Green) |
| Catalog No.GC26409 |
Lysotracker probes are acidotropic fluorescent probes used for labeling and tracking acidic organelles in living cells.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 215247-93-1
Sample solution is provided at 25 µL, 10mM.
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Lysotracker probes are acidotropic fluorescent probes used for labeling and tracking acidic organelles in living cells. These probes have several important features, including highly selective targeting of acidic organelles and efficient labeling of living cells at nanomolar concentrations. In addition, the LysoTracker series is available in several fluorescent colors, making it particularly suitable for multicolor labeling experiments.
Lysotracker probes, which contain a fluorescein linked to a weak base, are partially protonated at neutral pH, can freely penetrate the cell membrane and typically accumulate on spherical organelles. Studies have shown that Lysotracker probes must be used at very low concentrations (usually around 50nM) to achieve excellent selectivity. The retention mechanism of these probes, although not fully understood, is likely related to protonation and retention in acidic organelles, even though subsequent treatment of cells with weakly alkaline cell permeable compounds does not usually reverse the staining results. Studies on the internalization kinetics of Lysotracker probes have shown that the uptake rate of the dye into living cells is only a few seconds. However, these lysosomal probes can cause lysosomes to become alkaline, such as long-term incubation that induces an increase in lysosomal pH. Therefore, it is recommended to incubate cells with the probe for only 1-5 minutes before imaging.
LysoTracker Blue DND-22 is a blue fluorescent dye that stains acidic compartments in living cells, with a maximum excitation/emission wavelength of ~373/422nm. This product is stored in DMSO at a concentration of 1mM. For lysosomal staining of living cells, the recommended working concentration is 50-75nM, which needs to be optimized and adjusted according to actual conditions.
Precautions
1) Avoid light during the entire operation.
2) For your safety and health, please wear a lab coat and disposable gloves.
Usage
Before use, take out the product and warm it to room temperature, and centrifuge it at a low speed to make the solution drop to the bottom of the tube. For the first use, please store the product in a single-use aliquot, ≤-20℃ away from light and dry, and try to avoid repeated freezing and thawing. The optimal working concentration needs to be optimized according to different experimental requirements, cell types, and membrane permeability of cells or tissues.
1. Preparation of working solution
Use growth medium or appropriate buffer to dilute the 1mM storage solution to the working concentration. The working solution must be prepared and used immediately. For Lysotracker series lysosomal probes, the recommended working solution concentration is 50-75nM;
【Note】:
1) In order to reduce the false positives that may be caused by excessive probe loading, it is recommended to use as low a concentration as possible without affecting the staining effect.
2) If the cells are incubated in a dye-free medium after staining, the attenuation of the fluorescence signal and the vacuolization of the cells will be observed.
2. Staining steps (adherent cells)
1) Place the cells on a coverslip in a culture dish and add appropriate culture medium to allow them to grow on the slide.
2) When the cells grow to an appropriate density, remove the culture medium and add an appropriate amount of probe-containing culture medium preheated at 37°C (the cells must be completely covered). Incubate the cells for 30 min-2 h under growth conditions suitable for the specific cell type (the specific incubation time depends on the cell type).
3) Replace the above staining solution with fresh culture medium without probes and observe with a fluorescence microscope equipped with a suitable filter (see Appendix 1). If the staining is not sufficient, it is recommended to increase the dye concentration or extend the staining time so that the dye can accumulate on the lysosomes.
3. Staining steps (suspended cells)
1) Centrifuge the sedimented cells and remove the supernatant.
2) Resuspend the cells with an appropriate amount of probe-containing culture medium preheated at 37°C and incubate the cells for 30 min-2 h under growth conditions suitable for the specific cell type (the specific incubation time depends on the cell type).
3) Centrifuge, remove the staining solution, and add fresh culture medium to resuspend the cells.
4) Observe with a fluorescence microscope equipped with appropriate filters. If the staining is not sufficient, it is recommended to increase the dye concentration or extend the staining time so that the dye can accumulate on the lysosomes.
| Cas No. | 215247-93-1 | SDF | |
| Synonyms | LysoTracker Blue DND-22; LysoTracker Red DND-99; Lysotracker Green | ||
| Formula | C24H38Cl4N4 | M.Wt | 524.403 |
| Solubility | Storage | Desiccate at -20°C ; protect from light | |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.9069 mL | 9.5347 mL | 19.0693 mL |
| 5 mM | 0.3814 mL | 1.9069 mL | 3.8139 mL |
| 10 mM | 0.1907 mL | 0.9535 mL | 1.9069 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
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Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)
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