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| Targets |
2,6‑Diiodo‑BODIPY 493/503 does not target a specific protein; instead, its lipophilic nature allows it to partition into lipid‑rich cellular environments, including lipid droplets, endoplasmic reticulum membranes, and plasma membranes. The dye intercalates into the hydrophobic bilayer or neutral lipid core of droplets, driven by van der Waals interactions. Its preferential accumulation in lipid droplets makes it a specific marker for studying adipocyte differentiation, lipid metabolism, and neutral lipid storage disorders. The iodine atoms also confer singlet oxygen generation properties, which can be exploited for photodynamic therapy applications, though this is secondary to its staining utility.
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| ln Vitro |
In vitro, 2,6‑Diiodo‑BODIPY 493/503 is used to visualise and quantify lipid droplets in cultured mammalian cells, such as 3T3‑L1 adipocytes, HeLa cells, or primary hepatocytes. The dye is added to live cells at concentrations of 0.1‑1 µM for 15‑30 min; it rapidly labels neutral lipids with high specificity and minimal background fluorescence. Staining is independent of cell fixation, allowing live‑cell time‑lapse imaging. The dye exhibits no detectable cytotoxicity at working concentrations, as assessed by LDH release or propidium iodide exclusion. It can be multiplexed with other fluorophores (e.g., DAPI, phalloidin) due to its narrow emission spectrum.
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| ln Vivo |
In vivo, 2,6‑Diiodo‑BODIPY 493/503 has been applied in zebrafish and mouse models to study lipid metabolism and adipose tissue distribution. In zebrafish larvae, the dye is added to the water (1 µM) for 30 min; it specifically stains visceral and subcutaneous fat depots, enabling real‑time visualisation of lipogenesis and lipolysis upon dietary manipulation. In mice, the dye can be administered via tail‑vein injection (0.1‑0.5 mg/kg) to label circulating lipoproteins and adipose tissue; however, its use is mostly limited to ex vivo tissue staining of frozen sections due to rapid clearance and metabolic modification.
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| Enzyme Assay |
In vitro staining protocols are straightforward: cells grown on coverslips or in 96‑well plates are washed with PBS, then incubated with 0.5‑1 µM dye in serum‑free medium or HBSS for 15‑30 min at 37°C. The cells are washed twice with PBS and immediately imaged (ex/em ~493/503 nm) or fixed with 4% paraformaldehyde for later analysis. For quantification, stained cells are trypsinised, and fluorescence intensity is measured by flow cytometry (FITC channel). Competitive displacement experiments with other lipophilic dyes (e.g., Nile Red) can be performed to assess specificity. No pre‑treatment with fixing agents is needed for live cells.
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| Cell Assay |
Cell‑based assays often involve lipid droplet induction: cells are cultured with 0.5 mM oleic acid‑BSA complex for 24‑48 h to stimulate lipid accumulation. After treatment, 2,6‑Diiodo‑BODIPY 493/503 staining is performed as described above. The number and size of lipid droplets can be quantified using ImageJ or high‑content imaging software. For photodynamic studies, cells are incubated with the dye (1‑5 µM) for 1 h and then exposed to green light (532 nm) at 10‑50 J/cm²; cell viability is measured by MTT to assess light‑induced cytotoxicity due to singlet oxygen generation.
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| Animal Protocol |
In vivo protocols in zebrafish: embryos at 5‑10 dpf are placed in embryo water containing 0.5‑1 µM dye for 30 min, then rinsed and anaesthetised for imaging under a fluorescence stereomicroscope. For mouse studies, the dye is dissolved in DMSO and diluted in saline, then administered IV (0.5 mg/kg) or IP (1 mg/kg). After 1‑4 h, tissues (liver, adipose, muscle) are harvested, snap‑frozen, and cryosectioned (10 µm); sections are stained post‑sectioning with DAPI and imaged. Alternatively, the dye can be added to homogenised tissue for lipid extraction and fluorescence quantification.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of 2,6‑Diiodo‑BODIPY 493/503 are not extensively characterised, as it is a research dye. In rodents, after IV injection, the dye rapidly distributes to lipophilic compartments with a distribution half‑life of <5 min. It is cleared by hepatic uptake and biliary excretion, with a terminal half‑life of approximately 2‑4 h. The dye is highly lipophilic (logP > 5), leading to extensive binding to plasma lipoproteins and red blood cells. It is not metabolised to a significant extent; the intact dye is excreted in faeces. There is no data on human PK, as it is not a therapeutic agent.
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| Toxicity/Toxicokinetics |
Toxicological evaluation: 2,6‑Diiodo‑BODIPY 493/503 is generally considered non‑toxic at the low concentrations used for staining (0.1‑1 µM). In cell culture, no effects on proliferation or apoptosis are observed up to 10 µM. In zebrafish, continuous exposure at 5 µM for 24 h does not affect survival or morphology. The iodine substituents increase the risk of phototoxicity upon intense illumination, but this is minimal under standard imaging conditions. The dye is not classified as hazardous for routine laboratory handling, though standard safety precautions (gloves, lab coat) are advised.
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| References | |
| Additional Infomation |
Additional information: 2,6‑Diiodo‑BODIPY 493/503 is a specialised fluorescent probe for lipid research, with CAS 1031443‑55‑6. Its spectral properties (absorption ~493 nm, emission ~503 nm) make it compatible with standard GFP/FITC filter sets. Compared to Nile Red, it has narrower emission and higher photostability, allowing better spectral unmixing. The compound is not intended for clinical or therapeutic use; it is solely a research chemical. It is commonly supplied as a crystalline solid and stored at –20°C protected from light. It is a valuable tool for exploring lipid metabolism in health and disease.
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| Molecular Formula |
C14H15BF2I2N2
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| Molecular Weight |
513.90
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| CAS # |
1031443-55-6
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| Appearance |
Brown to red solid powder
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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) |
Typically soluble in DMSO (e.g. 10 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 | 1.9459 mL | 9.7295 mL | 19.4590 mL | |
| 5 mM | 0.3892 mL | 1.9459 mL | 3.8918 mL | |
| 10 mM | 0.1946 mL | 0.9730 mL | 1.9459 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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