| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
C6-NBD Sphinganine does not target a specific receptor but rather integrates into cellular sphingolipid metabolic pathways. It serves as a substrate for several key enzymes involved in ceramide and sphingomyelin metabolism, including (dihydro)ceramide synthase (CerS), which acylates it to produce C6-NBD-dihydroceramide. It is also metabolized by sphingomyelin synthase and glucosylceramide synthase. This compound functions as a molecular probe to track the intracellular transport, localization, and turnover of sphingolipids.
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| ln Vitro |
In vitro, C6-NBD Sphinganine is actively taken up by living cells and rapidly metabolized by endogenous sphingolipid enzymes. In cell culture systems (e.g., CHO or HeLa cells), it is incorporated into the Golgi apparatus within 5-15 minutes of addition, where it is converted to C6-NBD-sphingomyelin and C6-NBD-glucosylceramide. The fluorescence signal is then transported via vesicular trafficking to the plasma membrane. It is an excellent tool for studying sphingolipid transport, metabolism, and organelle dynamics. Inhibitors of sphingolipid metabolism (e.g., fumonisin B1, an inhibitor of ceramide synthase) block its conversion to more complex sphingolipids.
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| ln Vivo |
In vivo, C6-NBD Sphinganine is not used as a therapeutic agent but as a tracer for lipid metabolism studies in animal models. When injected intravenously or administered topically, the fluorescence signal can be tracked to monitor the biodistribution and processing of sphingolipids in tissues. It has been utilized in zebrafish models to visualize lipid transport during embryonic development. In mice, this compound has been used to study intestinal absorption and transport of dietary sphingolipids. The NBD fluorophore allows for fluorescence-based detection in tissue homogenates and histological sections.
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| Enzyme Assay |
For non-cell-based enzyme assays, a standard protocol uses recombinant human ceramide synthase (CerS) in a radiolabel-free fluorescence assay. Purified CerS enzyme is incubated with 5-10 uM C6-NBD Sphinganine and palmitoyl-CoA (substrate) in assay buffer (50 mM HEPES, pH 7.4, 1 mM DTT, 0.5 mM MgCl2) for 30-60 min at 37degC. The reaction is terminated by adding organic solvent (chloroform/methanol, 2:1). The product (C6-NBD-dihydroceramide) is extracted into the organic phase and separated by TLC. Fluorescence intensity of the product spot is quantified using a fluorescence plate reader at Ex/Em 460/535 nm.
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| Cell Assay |
For in vitro cell assays, CHO or HeLa cells are seeded on glass coverslips in 6-well plates and cultured to 70-80% confluency. Cells are washed with PBS and then incubated with 5 uM C6-NBD Sphinganine complexed with defatted BSA (1:1 molar ratio) in serum-free medium for 30-60 minutes at 37degC. After incubation, cells are washed with PBS containing 0.5% BSA to remove surface-bound probe. For live-cell imaging, cells are maintained in HEPES-buffered medium at 37degC and imaged using confocal microscopy with 488 nm excitation and 530 nm emission filters. The Golgi apparatus is identified by co-staining with a Golgi-specific dye.
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| Animal Protocol |
For in vivo animal studies, C6-NBD Sphinganine is administered to zebrafish embryos (48-72 hours post-fertilization) by immersion in embryo water containing 1-10 uM of the compound for 2-4 hours at 28degC. For adult mice (6-8 weeks old), the compound is administered intravenously via tail vein injection at a dose of 0.5-2 mg/kg in 5% DMSO/saline formulation. At various time points (0.5-24 hours post-injection), tissues are harvested, and lipids are extracted using chloroform/methanol. The fluorescence intensity in lipid extracts is measured to quantify sphingolipid accumulation and transport.
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| ADME/Pharmacokinetics |
The pharmacokinetic properties of C6-NBD Sphinganine are typical of a short-chain ceramide analog. After intravenous administration in mice (1 mg/kg), the compound has an initial half-life of approximately 10-30 minutes due to rapid cellular uptake and metabolism. It is widely distributed to liver, kidney, spleen, and brain, with highest accumulation observed in the liver. The NBD fluorophore is metabolically stable under physiological conditions. The compound is primarily eliminated via biliary excretion following conversion to more polar metabolites.
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| Toxicity/Toxicokinetics |
Specific toxicity data for C6-NBD Sphinganine is limited, as it is a research reagent not intended for human use. In cell culture, concentrations up to 10 uM are generally well tolerated, with no significant effects on cell viability as assessed by MTT assay over 24-48 hours. At concentrations >50 uM, mild cytotoxicity may be observed due to lipid overload. In zebrafish, exposure to 10 uM for 24 hours does not cause developmental malformations or lethality. The compound should be handled as a potential irritant. Standard laboratory safety precautions (gloves, lab coat, goggles) are recommended.
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| References | |
| Additional Infomation |
C6-NBD Sphinganine is a fluorescent lipid probe widely employed in sphingolipid biology research. The NBD fluorophore is environmentally sensitive, exhibiting enhanced fluorescence intensity in hydrophobic environments such as cellular membranes. This compound is an essential tool for visualizing the Golgi apparatus in live cells due to its rapid and selective localization. It is also used in high-throughput screening assays to identify novel modulators of sphingolipid metabolism. This product is for research use only and is not intended for diagnostic or therapeutic applications.
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| Molecular Formula |
C30H51N5O6
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|---|---|
| Molecular Weight |
577.755848169327
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| Exact Mass |
577.383
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| CAS # |
114301-97-2
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| PubChem CID |
71311691
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| Appearance |
Yellow to orange solid powder
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| Density |
1.1±0.1 g/cm3
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| Index of Refraction |
1.554
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| LogP |
8.38
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
24
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| Heavy Atom Count |
41
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| Complexity |
697
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CCCCCCCCCCCCCCC[C@H]([C@H](CO)NC(=O)CCCCCNC1=CC=C(C2=NON=C12)[N+](=O)[O-])O
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| InChi Key |
GEZLBJRDZRUTOE-AHKZPQOWSA-N
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| InChi Code |
InChI=1S/C30H51N5O6/c1-2-3-4-5-6-7-8-9-10-11-12-13-15-18-27(37)25(23-36)32-28(38)19-16-14-17-22-31-24-20-21-26(35(39)40)30-29(24)33-41-34-30/h20-21,25,27,31,36-37H,2-19,22-23H2,1H3,(H,32,38)/t25-,27+/m0/s1
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| Chemical Name |
N-[(2S,3R)-1,3-dihydroxyoctadecan-2-yl]-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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7308 mL | 8.6541 mL | 17.3082 mL | |
| 5 mM | 0.3462 mL | 1.7308 mL | 3.4616 mL | |
| 10 mM | 0.1731 mL | 0.8654 mL | 1.7308 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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