BODIPY-505/515 does not have a specific biological target in the context of a drug, but rather it is a fluorescent probe used to visualize and track cellular components. It localizes to intracellular lipid bodies and is used to label lipid droplets in live and fixed cells. Its mechanism of action is purely biophysical: upon excitation with light, the molecule emits fluorescence that can be detected and quantified. This property allows researchers to study lipid metabolism, cellular trafficking, and membrane dynamics without interfering with normal cellular processes.

Guidelines for use 1. Working solution for BODIPY505/515 stock solution 1.1 Get the stock solution ready. 382 μL of anhydrous DMSO should be loaded with 1 milligram of BODIPY505/515. Take out the 10 mM reference solution. Note: It is advised to prepare a 1–10 μM BODIPY505/515 working solution by either aliquoting the BODIPY505/515 storage solution at -20°C or using pre-made serum-free cell culture medium or PBS filling solution with the 1.2 working solution. Note: Please keep BODIPY505 at -80°C away from light and adjust to real conditions. 2. Cell staining (suspended cells) 2.1 Centrifuge the cells, add PBS, and wash twice for five minutes each time. Following a centrifugation for 3–4 minutes and the removal of the supernatant, add 1 mL BODIPY505/515 2.3 400 g when the cell density reaches 1×106/mL 2.2. 2.4 Wash the cells twice, for five minutes each time, with PBS added. 2.5 Resuspend in 1 milliliter of PBS 3 or serum-free water. Staining of adherent cells 3.1 The adherent cells are cultured on sterile coverslips. 3.2 Aspirate extra culture after removing the coverslip from the cells. 3.3 After adding 100 μL of the dye working solution, fully cover the cells with your hand waves. It will require between five and thirty minutes to complete. 3.4 Aspirate the dye working solution, use a fluorescence microscope or base, and wash it twice, for a duration of five minutes each, in culture media. using flow cytometry observation. Conditions for storage: Keep away from light and store at -20°C for a year. Notes 1. Kindly modify the BODIPY505/515 working fluid concentration and wastewater time to reflect the current circumstances. 2. In the experiment, it is advised to use laboratory control. Before doing following tests, incubate the document cells with the cells in a gradient of 30 μM oleic acid for 8 hours. 3. This product may not be used in food or medication, nor may it be utilized for clinical diagnosis or treatment. It is intended solely for professional use in scientific study. 4. Please use a lab coat and disposable gloves for your health and safety.
In vitro, BODIPY-505/515 is used as a fluorescent staining dye for lipid membranes. It is very bright and possesses excellent stability under excitation irradiation, making it ideal for long-term imaging experiments. The dye is used to generate fluorescent conjugates of proteins and to prepare fluorescent enzyme substrates, fatty acids, receptor ligands, and polystyrene microspheres. Its fluorescence emission in the green to yellow-green region makes it useful for multiplexed imaging and detection in various assays, including flow cytometry, live cell imaging, and fluorescence microscopy.

In vivo activity data for BODIPY-505/515 is not typically reported, as it is a fluorescent probe rather than a bioactive drug. It is primarily used in vitro or ex vivo for labeling and imaging purposes. However, the dye can be used to label cells or biomolecules that are subsequently administered to animals for tracking purposes. For example, BODIPY-labeled cells can be injected into mice to study cell migration or tumor homing. In such applications, the dye serves as a tracer, and its fluorescence can be detected in tissues using imaging systems.

In vitro enzyme/receptor binding (non-cell) assays using BODIPY-505/515 typically involve the dye as a fluorescent label rather than as a binding agent itself. A common protocol for labeling proteins or peptides involves dissolving BODIPY-505/515 in an appropriate organic solvent (e.g., DMSO or DMF) and reacting it with the target biomolecule under mild conditions. The reaction mixture is incubated at room temperature for 1-2 hours, and the labeled product is purified by size exclusion chromatography or dialysis. The degree of labeling is determined by measuring the absorbance and fluorescence of the conjugate. This approach is used to create fluorescently labeled ligands for receptor binding studies.

In vitro cell-based assays for BODIPY-505/515 involve staining cells to visualize specific structures. A standard protocol for lipid droplet staining: cells are grown on coverslips or in a 96-well plate. The culture medium is removed, and cells are washed with PBS. A solution of BODIPY-505/515 (typically 1-10 µM) in PBS or serum-free medium is added to the cells, followed by incubation at 37°C for 15-30 minutes. Cells are then washed to remove unbound dye. Fluorescence is visualized using a fluorescence microscope with appropriate excitation and emission filters (excitation ~505 nm, emission ~515 nm). The dye can also be used for flow cytometry analysis.

In vivo animal experiments using BODIPY-505/515 are typically not performed for the dye itself, but rather for BODIPY-labeled biomolecules or cells. A standard protocol for tracking labeled cells: cells are stained with BODIPY-505/515 in vitro, washed, and resuspended in PBS. The labeled cells (e.g., 1×10⁶ cells) are then injected intravenously or subcutaneously into immunocompromised mice. At various time points post-injection, animals are euthanized, and tissues are collected. Tissue sections are prepared and analyzed under a fluorescence microscope to detect the BODIPY signal, allowing researchers to track the distribution and migration of the labeled cells.

Pharmacokinetic properties of BODIPY-505/515 are not a primary focus of study, as it is a research tool rather than a therapeutic agent. The compound is highly lipophilic, which contributes to its ability to partition into lipid membranes and lipid droplets. It is generally considered to be cell-permeable due to its lipophilic nature. In biological systems, the dye is typically retained within cells for extended periods, making it suitable for long-term imaging experiments. Its stability under various physiological conditions, including different pH and polarity environments, is a key advantage for its use as a fluorescent probe.

Toxicity data for BODIPY-505/515 is limited. As a fluorescent dye, it is generally considered to have low toxicity at the concentrations used for cell labeling and imaging (typically in the micromolar range). BODIPY dyes are known for their excellent stability and low photobleaching, and they are generally well-tolerated by cells. However, as with all chemical reagents, appropriate safety precautions should be taken when handling the compound. It should be stored protected from light and handled with standard laboratory safety equipment.

[1]. Detection and quantitation of lipid in the microalga Tetraselmis subcordiformis (Wille) Butcher with BODIPY 505/515 staining. Bioresour Technol. 2013 Jan;127:386-90.

[2]. BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry. Bio Protoc. 2016 Sep 5;6(17):e1912.

BODIPY-505/515 is a widely used fluorescent probe in molecular biology and biochemistry. It belongs to the BODIPY class of dyes, which are characterized by strong UV absorption, sharp fluorescence peaks, and high quantum yields. The dye's insensitivity to environmental polarity and pH makes it a robust tool for various physiological conditions. Its applications include lipid droplet labeling, flow cytometry, live cell imaging, and fluorescence microscopy. It is also used to generate fluorescent conjugates of proteins, enzyme substrates, and receptor ligands. The compound is not a drug and has no clinical use or approval status. It is commercially available from chemical suppliers for research purposes only.

C13H15BF2N2

248.0794

248.129

21658-70-8

16679984

Brown to red solid powder

0

3

0

18

468

0

F[B-]1(N2C(C([H])([H])[H])=C([H])C(C([H])([H])[H])=C2C([H])=C2C(C([H])([H])[H])=C([H])C(C([H])([H])[H])=[N+]21)F

TWYZTUZOEQTCOO-UHFFFAOYSA-N

InChI=1S/C13H15BF2N2/c1-8-5-10(3)17-12(8)7-13-9(2)6-11(4)18(13)14(17,15)16/h5-7H,1-4H3

2,2-difluoro-4,6,10,12-tetramethyl-3-aza-1-azonia-2-boranuidatricyclo[7.3.0.03,7]dodeca-1(12),4,6,8,10-pentaene

2934.99.9001

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.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~12.5 mg/mL (~50.39 mM)

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)