| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
The probe targets acidic cellular compartments (specifically lysosomes). Its mechanism of action is based on a physicochemical property rather than a biological receptor. The weak base moiety becomes protonated within the acidic lysosomal environment (pH ~4.5-5.0), which prevents it from diffusing back across the membrane, leading to selective accumulation of the fluorescent dye. LysoTracker probes are not fixable, so they must be used on live cells.
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| ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular circumstances as it simply offers guidance). Quantitative GAPDH LysoTracker Yellow HCK 123 Assay [1]: 1. Utilizing 8-well cell culture slides, cultivate EMT6 cells. Use a standard methodology for incubating cells. 2. To get the final concentration of 30 µg/mL, add TONS504 to each well. For four hours, incubate the cells at 37 °C. 3. To each well, add LysoTracker Yellow HCK 123 until the final concentration reaches 50 μM. 4. The cells were incubated for 30 minutes at 37°C with 5% CO2. Use a confocal laser scanning microscope to examine samples.
As a fluorescent probe for imaging, it does not exert a pharmacological "activity" like a drug. Its "activity" is its ability to fluoresce when excited (Ex=465 nm) and accumulate in lysosomes. The fluorescence intensity is directly proportional to the acidity of the organelles, making it a useful tool for studying lysosomal pH dynamics, autophagic flux, and other cellular processes involving acidic vesicles. |
| ln Vivo |
LysoTracker is an in vitro diagnostic imaging probe and is not used for in vivo "activity" in the context of a drug. Therefore, it does not have in vivo efficacy data like a therapeutic. It is used in pre-clinical in vitro assays to study lysosomal biology and can be used as a marker for autophagy or endocytosis in cells derived from animal tissues.
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| Enzyme Assay |
This is a fluorescent dye for cell staining; it is not an enzyme inhibitor and is not used in cell-free enzyme binding assays. A cell-free application is not applicable. Its selectivity is based on the cell's intact physiology, not a purified protein.
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| Cell Assay |
The standard protocol for LysoTracker Yellow HCK 123 is as follows: 1. Prepare a 1 mM stock solution in DMSO. 2. Dilute the stock solution to the working concentration (typically 50-75 nM) in warm, serum-free cell culture medium. 3. For live cells, aspirate the culture medium and add the pre-warmed LysoTracker-containing medium. 4. Incubate cells for 30-120 minutes at 37degC in a CO2 incubator. 5. Wash the cells gently with pre-warmed PBS or medium 2-3 times. 6. Image live cells immediately using a fluorescence microscope or confocal with a filter set for yellow fluorescence (Ex/Em ~465/535 nm).
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| Animal Protocol |
LysoTracker Yellow HCK 123 is strictly an in vitro live-cell imaging probe. Typical cell-based experiments for lysosomal tracking involve: 1. Seed cells (e.g., HeLa, EMT6) into 8-well chamber slides or 96-well imaging plates and allow to adhere. 2. Perform any cellular treatments (e.g., drug stimulation, starvation) as per the experiment's design. 3. Stain live cells with LysoTracker (50-75 nM) for 30-120 min at 37degC in the dark. 4. Optionally, counterstain the nucleus with Hoechst 33342. 5. Wash and replace with fresh medium, then image immediately. The probe cannot be used on fixed/permeabilized cells.
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| ADME/Pharmacokinetics |
LysoTracker Yellow HCK 123 is not designed for in vivo animal experiments due to high background and metabolism. While some analogous probes are used for intravital imaging, this specific product is typically for in vitro use. An in vivo procedure would involve tail vein injection and then whole-body fluorescence imaging, but the utility is limited due to rapid clearance and non-specific accumulation.
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| Toxicity/Toxicokinetics |
Not applicable. This is an imaging dye, not a systemic drug. Pharmacokinetic properties are not relevant as it is used directly on cells in culture for immediate imaging. Its properties are defined by its spectral characteristics (λex=465 nm, λem=535 nm), cell permeability, and lysosomal retention.
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| References | |
| Additional Infomation |
HCK-123 dye is a benzoxadiazole compound that functions as a fluorescent dye.
Not applicable (non-toxic fluorescent dye at standard working concentrations). The compound is a weakly basic amine that is generally considered non-toxic to cells at the recommended nanomolar concentrations. Its mechanism relies on partitioning into a subcellular compartment, not on inducing cytotoxicity, and it is used for short-term live-cell imaging experiments. |
| Molecular Formula |
C16H24N6O4
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|---|---|
| Molecular Weight |
364.4
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| Exact Mass |
364.185
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| CAS # |
1064123-31-4
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| PubChem CID |
25195435
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| Appearance |
Yellow to orange solid powder
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| LogP |
1.3
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
10
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| Heavy Atom Count |
26
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| Complexity |
459
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| Defined Atom Stereocenter Count |
0
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| SMILES |
N(C1=CC=C(N(=O)=O)C2=NON=C12)CCCCCC(=O)NCCN(C)C
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| InChi Key |
WIOHDRVBBRICQL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C16H24N6O4/c1-21(2)11-10-18-14(23)6-4-3-5-9-17-12-7-8-13(22(24)25)16-15(12)19-26-20-16/h7-8,17H,3-6,9-11H2,1-2H3,(H,18,23)
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| Chemical Name |
N-[2-(dimethylamino)ethyl]-6-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]hexanamide
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~62.5 mg/mL (~171.51 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.71 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7442 mL | 13.7212 mL | 27.4424 mL | |
| 5 mM | 0.5488 mL | 2.7442 mL | 5.4885 mL | |
| 10 mM | 0.2744 mL | 1.3721 mL | 2.7442 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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