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| Product Name | Lysosomal pH Green Fluorescent Probe (DND-189) |
| Catalog No. | RESK-0558 |
| Description | This series of probes comprises fluorescent probes for studying the dynamic synthesis and function of acidic compartments in living cells, such as lysosomes and sperm acrosomes. The probes are acidotropic: they accumulate in acidic organelles through protonation. This protonation also relieves fluorescence quenching of the dye, resulting in stronger emitted fluorescence. Therefore, unlike pH-independent lysosomal probes (such as the DND-26/DND-99 type), these pH-sensitive probes show a pH-dependent (acid-directed) increase in fluorescence intensity as the organelle acidifies. The Lysosomal pH Green Fluorescent Probe (DND-189) has a relatively low pKa value and is fluorescent only within acidic compartments. It is supplied as a 1 mM stock solution in anhydrous DMSO. For selective labeling and staining of acidic compartments in live cells independent of pH, pH-independent lysosomal probes may be chosen instead. |
| Intended Use | pH-dependent fluorescent labeling and study of acidic compartments (e.g., lysosomes, sperm acrosomes) in living cells, including monitoring of organelle acidification. |
| Principle / Technology | The acidotropic probe accumulates in acidic organelles via protonation. Protonation relieves fluorescence quenching, so fluorescence intensity increases in a pH-dependent manner as the compartment acidifies. With a pKa of approximately 5.2, DND-189 is essentially fluorescent only inside acidic compartments. |
| Detection Method | Green fluorescence detection (Ex/Em 443 nm/505 nm) by fluorescence microscope, confocal microscope, microplate reader, or flow cytometer. |
| Sample Type | Living cells only; staining must be performed while cells are alive. Plant cells or tissues are theoretically applicable and generally need to be prepared as protoplasts. |
| Performance Range / Specifications | Molecular formula: C24H22N4O2 Molecular weight: 398.46 Ex/Em: 443/505 nm pKa: ~5.2 Color: green Applicable pH detection range: 4.5-6.0 |
| Specificity | pH-dependent probe for acidic compartments in live cells; primarily intended for staining conventional lysosomes. Staining performance on autolysosomes is uncertain because autolysosomes differ considerably from normal lysosomes in morphology and structure; published literature may be consulted regarding staining of other acidic organelles. |
| Reaction Conditions / Protocol | Stain living cells; prepare the working solution fresh in cell culture medium immediately before use and do not store it. Perform staining protected from light, and detect the fluorescent signal immediately after staining, as the signal gradually weakens over time; do not leave stained samples overnight before detection. Excessively high probe concentration or overly long staining time (e.g., 50 μM for 50 min) can cause fluorescent signal in the cytoplasm in addition to lysosomes; reduce the concentration or shorten the staining time if this occurs. |
| Components / Formulation | Lysosomal pH Green Fluorescent Probe (DND-189), supplied as a 1 mM stock solution in anhydrous DMSO. |
| Storage Conditions | Store at -20 °C, protected from light and dry; avoid repeated freeze-thaw cycles. |
| Shelf Life | Six months when stored at -20 °C, protected from light. |
| Package Specifications | 50 μL |
| Product Form | Solution (1 mM stock in anhydrous DMSO) |
| Concentration | 1 mM stock solution in anhydrous DMSO |
| Molecular Weight | 398.46 |
| pH Range / Optimal pH | Applicable pH detection range: 4.5-6.0; pKa ~5.2 |
| Shipping Conditions | Shipped with ice packs. |
| Expiration Date / Stability | Valid for six months when stored at -20 °C, protected from light and dry; avoid repeated freeze-thaw cycles. Working solutions should be prepared fresh and are not recommended for storage. |
| Regulatory / Compliance | For research use only. |
| Compatibility | For simultaneous staining of lysosomes and nuclei, Hoechst 33258 or Hoechst 33342 is recommended for nuclear staining, as these dyes stain live cells directly without fixation. Cannot be co-incubated with secondary antibodies, because the permeabilization required for antibody incubation destroys the acidic lysosomal environment and impairs staining. |
| Recommended Buffer System | Prepare the working solution fresh in cell culture medium; use immediately and do not store. |
| Application Notes / Precautions | Do not fix cells before or after staining: fixation with paraformaldehyde, aldehydes, etc. disrupts the acidic lysosomal environment and can weaken or abolish fluorescence. Do not permeabilize cells; permeabilization likewise disrupts lysosomal acidity. Protect from light during staining and minimize the interval between staining and detection. If the signal is weak: appropriately increase the probe concentration, extend the staining time, and detect immediately after staining. If cytoplasmic background appears: reduce the probe concentration or shorten the staining time. |
For research use only, not for clinical use.
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