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| Product Name | Lysosome Red Fluorescent Probe (DND-99) |
| Catalog No. | RESK-0545 |
| Description | This probe belongs to a family of fluorescent dyes that selectively stain acidic compartments in live cells. These probes offer several important features: 1) selective labeling of acidic organelles; 2) effective labeling of live cells at nanomolar (nM) concentrations; 3) availability in multiple colors, allowing multicolor labeling experiments as needed. Structurally, the probe consists of a fluorophore linked to a weak base. It freely crosses cell membranes and typically accumulates in spherical organelles, making it suitable for studying lysosome biogenesis and related pathogenesis. At neutral pH the probe is only partially protonated, so its organelle-labeling mechanism is thought to involve full protonation and retention on organelle membranes. Lysosome Red Fluorescent Probe (DND-99) is the red fluorescent member of this family, with excitation/emission maxima of 577/590 nm. It is supplied as a 1 mM stock solution in anhydrous DMSO. |
| Intended Use | Selective fluorescent staining of acidic organelles (lysosomes) in live cells for fluorescence imaging and analysis of lysosomal biogenesis and related pathogenic mechanisms. |
| Principle / Technology | The probe is an acidotropic dye composed of a fluorophore linked to a weak base. It freely diffuses across cell membranes and, being only partially protonated at neutral pH, becomes fully protonated in acidic compartments and is retained on organelle membranes, resulting in selective accumulation in acidic organelles such as lysosomes. |
| Detection Method | Fluorescence microscopy; confocal microscopy; microplate reader; flow cytometry |
| Sample Type | Live cells (staining must be performed on live cells). Plant cells or tissues are theoretically applicable and are generally prepared as protoplasts first. |
| Performance Range / Specifications | Molecular formula: C20H24BF2N5O Molecular weight: 399.2493 Ex/Em: 577/590 nm Appearance: purple solution Concentration: 1 mM stock in anhydrous DMSO CAS No.: N/A |
| Sensitivity / LOD | Effective labeling of live cells at nanomolar (nM) concentrations; very low probe concentrations are recommended for preferential staining of acidic lysosomes. |
| Specificity | Selectively labels acidic organelles in live cells. Primarily intended for conventional lysosomes; staining of autolysosomes is not well characterized, and other acidic compartments may be labeled, as reported in the literature. |
| Reaction Conditions / Protocol | Dilute the 1 mM stock into prewarmed cell culture medium to a nanomolar working concentration immediately before use; the working solution is not stable for storage. Incubate live cells, then detect the fluorescence signal immediately. Avoid excessively high probe concentrations or overly long staining times, which cause non-specific cytoplasmic background fluorescence. |
| Components / Formulation | 1 mM stock solution of the red fluorescent lysosome probe (DND-99) in anhydrous DMSO |
| Storage Conditions | Store at -20°C, protected from light; avoid repeated freeze-thaw cycles. |
| Shelf Life | 1 year when stored at -20°C, protected from light. |
| Package Specifications | 50 μL |
| Product Form | Purple solution (1 mM in anhydrous DMSO) |
| Concentration | 1 mM stock solution |
| Molecular Weight | 399.2493 |
| Shipping Conditions | Shipped at room temperature. |
| Expiration Date / Stability | Valid for 1 year at -20°C, protected from light; avoid repeated freeze-thaw cycles. Working solutions prepared in cell culture medium should be used immediately and are not recommended for storage. |
| Regulatory / Compliance | For research use only. |
| Compatibility | Can be combined with Hoechst 33258 or Hoechst 33342 for simultaneous staining of lysosomes and nuclei in live cells, as these nuclear dyes stain live cells directly without fixation. Not compatible with fixation or permeabilization procedures (e.g., co-incubation with secondary antibodies), which destroy the acidic lysosomal environment. |
| Recommended Buffer System | Dilute into prewarmed cell culture medium for the working solution; prepare fresh before each use. |
| Application Notes / Precautions | Staining must be performed on live cells. Do not fix cells before or after staining (e.g., with paraformaldehyde or glutaraldehyde), and do not permeabilize, as this disrupts the acidic lysosomal environment and weakens or abolishes the signal. Protect from light during staining and detect fluorescence immediately after staining; signal fades over time. Prepare working solutions fresh; do not store them. Weak signal may result from too low a probe concentration, too short a staining time, or delayed detection after staining; increase concentration, extend staining, or detect immediately as appropriate. Cytoplasmic background in addition to lysosomal staining usually indicates excessive probe concentration or overly long staining; reduce the concentration or shorten the incubation. |
| Recognition Site | Acidic organelles (lysosomes) in live cells |
For research use only, not for clinical use.
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