| Size | Price | Stock | Qty |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
Nile Red does not have a pharmacological target but serves as a fluorescent probe for lipids and hydrophobic environments. It selectively stains neutral lipids, including triglycerides and cholesteryl esters, within cells. It can also label the hydrophobic domains of proteins and identify lysosomal phospholipid inclusions. Its fluorescence intensity and wavelength are sensitive to the polarity of the local environment.
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| ln Vitro |
1.1 Prepare the stock solution using DMSO to generate a 1mM stock solution. 1.2 Replace the working solution with warmed serum-free cell culture medium or PBS loading storage solution, with a final concentration of 200-1000 nM. Note: Please change the Nile Red work according to the actual situation. 2. Cell staining 2.1 Suspension cells: Collect cells by centrifugation, add PBS to wash twice, 5 minutes each time. Adherent cells: Discard the culture medium and add islet digested cells. After centrifugation and discarding the supernatant, add PBS and wash twice, 5 minutes each time. 2.2 Add 1 mL Nile Red working solution and proceed with filtration for 5-10 minutes. 2.3 Centrifuge at 400 g for 3-4 minutes at 4°C and discard the supernatant. 2.4 Wash the cells twice with PBS, giving them five minutes each time. 2.5 Resuspend the cells in 1 mL of serum-free or PBS and view using a fluorescence microscope.
In vitro, Nile Red is used as a fluorescent probe for intracellular lipids and hydrophobic domains of proteins. It selectively stains lipid droplets in a variety of cell types, making it a useful tool for studying lipid metabolism, adipocyte differentiation, and lipid-related diseases. Its fluorescence is quenched in aqueous environments but becomes strongly fluorescent in hydrophobic environments. Nile Red is also used to stain lysosomal phospholipid inclusions. |
| ln Vivo |
Looking at the green fluorescence of C stained with Nile red. elegans, distinct liposomes can be seen in the gut and other tissues, either uniformly distributed or in clusters, contingent upon the genotype of the animal and the manner in which the experiment was conducted[3].
In vivo, Nile Red is used as a fluorescent dye for imaging lipid-rich structures in live organisms, such as zebrafish larvae or C. elegans. It can be administered to live organisms to visualize lipid stores and fat metabolism. However, its utility is limited by its hydrophobicity and potential toxicity at high concentrations. |
| Enzyme Assay |
Nile Red is used in fluorescence microscopy and flow cytometry assays for lipid detection. Cells or tissues are incubated with Nile Red at concentrations typically ranging from 0.1 to 10 µg/mL in PBS or culture media for 5-30 minutes. After incubation, cells are washed and fluorescence is measured using a fluorescence microscope with excitation/emission of ~550-560/580-640 nm or by flow cytometry.
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| Cell Assay |
Nile Red is used to stain lipid droplets in cultured cells. Cells are seeded on coverslips or in multi-well plates, treated with Nile Red (0.1-10 µg/mL) for 5-30 minutes at 37°C, and then washed. Stained cells are visualized by fluorescence microscopy (excitation/emission ~550/580-640 nm). Lipid droplet content can be quantified by fluorescence intensity measurements or by image analysis.
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| Animal Protocol |
In vivo animal studies with Nile Red are conducted to visualize lipid stores in live organisms such as zebrafish larvae or C. elegans. Organisms are incubated with Nile Red in water or culture medium for 1-24 hours, washed, and then imaged using fluorescence microscopy. Lipid droplet distribution and abundance can be assessed in whole organisms or in specific tissues.
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| ADME/Pharmacokinetics |
Nile Red is a fluorescent dye and does not have pharmacokinetic properties as a drug. It is applied exogenously to cells or organisms in research settings. Its cellular uptake is driven by its lipophilic nature, and it accumulates in lipid-rich compartments. It is not administered systemically as a therapeutic agent.
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| Toxicity/Toxicokinetics |
Nile Red is a fluorescent dye and is not intended for human consumption or therapeutic use. It should be handled with appropriate laboratory safety precautions, including the use of gloves and eye protection. Some toxicity may occur at high concentrations in cell culture. It is classified as a potential irritant.
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| References |
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| Additional Infomation |
Nile red is an organoheterocyclic compound, a 5H-benzo[a]phenoxazine-5-one with a diethylamino group substituted at the 9-position. It can be used as a fluorescent dye and histological dye. It is an organoheterocyclic compound, a cyclic ketone, an aromatic amine, and a tertiary amine compound.
Nile Red is a polarity-sensitive fluorescent probe for neutral lipids and hydrophobic environments, widely used for staining intracellular lipid droplets and hydrophobic protein domains. It has applications in studying lipid metabolism, adipocyte differentiation, and lipid-related diseases. It is not a drug and has no clinical approval. |
| Molecular Formula |
C20H18N2O2
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|---|---|
| Molecular Weight |
318.36912
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| Exact Mass |
318.136
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| CAS # |
7385-67-3
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| PubChem CID |
65182
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| Appearance |
Green to dark green solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
484.7±45.0 °C at 760 mmHg
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| Melting Point |
203-205 °C(lit.)
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| Flash Point |
246.9±28.7 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
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| Index of Refraction |
1.644
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| LogP |
3.65
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
24
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| Complexity |
564
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| Defined Atom Stereocenter Count |
0
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| InChi Key |
VOFUROIFQGPCGE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C20H18N2O2/c1-3-22(4-2)13-9-10-16-18(11-13)24-19-12-17(23)14-7-5-6-8-15(14)20(19)21-16/h5-12H,3-4H2,1-2H3
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| Chemical Name |
9-(diethylamino)benzo[a]phenoxazin-5-one
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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) |
Ethanol : ~1 mg/mL (~3.14 mM)
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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 | 3.1410 mL | 15.7050 mL | 31.4100 mL | |
| 5 mM | 0.6282 mL | 3.1410 mL | 6.2820 mL | |
| 10 mM | 0.3141 mL | 1.5705 mL | 3.1410 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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