BODIPY borofluoroprene derivative
BODIPY 581/591 C11 targets lipid peroxidation products and reactive oxygen species (ROS) in cellular membranes. The probe is incorporated into cellular membranes, where it reacts with lipid peroxides and ROS generated during oxidative stress. Upon oxidation, the probe undergoes a spectral shift from red (591 nm) to green (510 nm), allowing the detection and quantification of lipid peroxidation. The compound is used as a ratiometric probe, where the ratio of green to red fluorescence provides a measure of the extent of lipid peroxidation. BODIPY 581/591 C11 is also used to detect ferroptosis, a form of iron-dependent cell death characterized by lipid peroxidation.

Preparation of stock solution:
The stock solution is prepared by dissolving 1 milligram of BODIPY 581/591 C11 in 0.1983 mL DMSO to produce 10 mM of the stock solution.
Note: Store the stock solution at -20°C or -80°C, protect from light, and avoid repeated freeze and thaw cycles.
1.2 Preparing the BODIPY 581/591 C11 working solutions:
Take the above BODIPY 581/591 C11 stock solution and dilute it to 2–10 μM with medium or PBS.
Note: You may adjust the concentrations of BODIPY 581/591 C11 working solution based on your experimental needs.
Cell staining:
2.1 Cell preparation:
Suspension cells: Centrifuge 1000g at 4 ℃ for 3-5 minutes and discard the supernatant. Wash twice with PBS for 5 minutes each time.
Adherent cells: Discard the cell culture medium, add trypsin to dissociate the cells, and prepare a single-cell suspension. Centrifuge at 1000 g at 4 ° C for 3-5 minutes, then discard the supernatant. Wash twice with PBS for 5 minutes each time.
2.2 Add 1 mL of BODIPY 581/591 C11 working solution and incubate at room temperature for 30 minutes.
2.3 Discard the supernatant after centrifuging at 400 g for three to four minutes at 4°C.
2.4 Use PBS twice, giving each wash five minutes.
2.5 Resuspend the cells for flow cytometry or fluorescence microscopy in PBS or serum-free cell culture media.
Note:
1. It is advised that the stock solution be stored at -20°C or -80°C away from light and avoid repeated freeze and thaw cycles.
2. You may adjust the concentrations of BODIPY 581/591 C11 working solution based on your experimental needs.
3. This product is not to be used in human; it is only to be used for research and development.
4. Please wear a white lab-coat and disposable gloves for your health and safety.
In vitro, BODIPY 581/591 C11 is used to detect lipid peroxidation in various cell types. The probe is incubated with cells at concentrations typically ranging from 1 to 10 μM for 30-60 minutes. After washing, cells are examined by fluorescence microscopy or flow cytometry. The red fluorescence (excitation 581 nm, emission 591 nm) is detected in the reduced (non-oxidized) state, while the green fluorescence (excitation 500 nm, emission 510 nm) appears upon oxidation. The ratio of green to red fluorescence provides a quantitative measure of lipid peroxidation. BODIPY 581/591 C11 has been used to study oxidative stress in various models, including neurodegenerative diseases, cardiovascular disease, and cancer.

In vivo, BODIPY 581/591 C11 can be used to detect lipid peroxidation in living organisms. The probe can be administered to animals via injection or other routes, and its fluorescence can be detected using imaging systems. BODIPY 581/591 C11 has been used to study oxidative stress in animal models of disease, including models of ischemia-reperfusion injury, neurodegeneration, and inflammation. The probe's ratiometric properties allow for semi-quantitative assessment of oxidative stress in tissues. However, the use of BODIPY 581/591 C11 in vivo is limited by its lipophilicity and the need for tissue penetration and clearance.

In vitro assays for BODIPY 581/591 C11 involve incubating cells or isolated membranes with the probe and measuring the fluorescence shift upon oxidation. Cells are seeded in culture plates and incubated with BODIPY 581/591 C11 at 2-10 μM for 30-60 minutes. After washing, cells are treated with oxidative stressors (e.g., H₂O₂, tert-butyl hydroperoxide, or iron) to induce lipid peroxidation. The fluorescence intensity at 591 nm (red) and 510 nm (green) is measured using a fluorescence microplate reader, flow cytometer, or fluorescence microscope. The ratio of green to red fluorescence is calculated as a measure of lipid peroxidation. The assay includes untreated controls and positive controls (cells treated with known oxidative stressors).

C11-BODIPY(581/591) is a fluorescent radio-probe for indexing lipid peroxidation and antioxidant efficacy in model membrane systems and living cells, with excellent characteristics: (i) emission in the visible range of the electromagnetic spectrum, with good spectral separation of the nonoxidized (595 nm) and oxidized (520 nm) forms; (ii) has a high quantum yield and because of this, low labeling concentrations can be used, ensuring minimal perturbation of the membrane whilst retaining favorable signal to noise ratios; (iii) has a good photo-stability and displays very few fluorescence artifacts; (iv) is virtually insensitive to environmental changes, i.e., pH or solvent polarity; (v) is lipophilic and as such easily enters membranes; (vi) once oxidized, C11-BODIPY(581/591) remains lipophilic and does not spontaneously leave the lipid bilayer; (vii) C11-BODIPY(581/591) localizes in two distinct pools within the lipid bilayer, a shallow pool at 18 A and a deep pool at < 7.5 A from the center of the bilayer; (viii) is not cytotoxic to rat-1 fibroblasts up to 50 microM; (ix) is sensitive to a variety of oxy-radicals and peroxynitrite, but not to superoxide, nitric oxide, transition metal ions, and hydroperoxides per se; (x) its sensitivity to oxidation is comparable to that of endogenous fatty acyl moieties.
In vitro cell-based assays for BODIPY 581/591 C11 are performed using various cell lines to study oxidative stress and lipid peroxidation. Cells are cultured in 96-well plates or on coverslips and incubated with BODIPY 581/591 C11 at 2-10 μM for 30-60 minutes. After washing, cells are treated with oxidative stressors or test compounds to induce or inhibit lipid peroxidation. The fluorescence is measured using a fluorescence microplate reader (excitation 488 nm, emission 510 nm and 590 nm) or by flow cytometry. The green/red fluorescence ratio is calculated. The effects of antioxidants or other compounds on lipid peroxidation can be assessed by comparing the fluorescence ratio in treated vs. untreated cells. Cell viability is assessed by MTT or other assays.

In vivo animal studies with BODIPY 581/591 C11 are conducted in rodent models of oxidative stress and disease. The probe is administered to animals via intravenous injection or other routes, and its distribution and oxidation are monitored by imaging. BODIPY 581/591 C11 has been used to study oxidative stress in models of ischemia-reperfusion injury, where the probe is injected prior to or after the induction of ischemia. Fluorescence imaging of tissues is performed to assess the extent of lipid peroxidation. However, detailed protocols for in vivo use of BODIPY 581/591 C11 are not extensively standardized, and the probe is primarily used in cell-based assays.

Pharmacokinetic properties of BODIPY 581/591 C11 are not typically characterized, as the compound is a fluorescent probe rather than a drug. The probe is lipophilic and is expected to partition into cellular membranes. Its distribution in vivo would depend on the route of administration and the lipophilicity of the compound. BODIPY 581/591 C11 is stable in the reduced state and becomes fluorescent upon oxidation. The probe's fluorescence properties allow for its detection in cells and tissues. For research use, the probe is typically stored at -20°C and protected from light.

BODIPY 581/591 C11 has low cytotoxicity at the concentrations typically used for fluorescence labeling (1-10 μM). The probe is well-tolerated by cells and does not significantly affect cell viability or function at these concentrations. No specific toxicological studies have been reported in the literature. As a research chemical, standard safety precautions should be followed when handling BODIPY 581/591 C11. The compound is for research use only and not for human or veterinary applications.

[1]. C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology. Free Radic Biol Med. 2002 Aug 15;33(4):473-90.

BODIPY 581/591 C11 is a widely used fluorescent probe for the detection of lipid peroxidation, a hallmark of oxidative stress and ferroptosis. Ferroptosis is a form of regulated cell death characterized by iron-dependent lipid peroxidation, which has been implicated in various diseases including cancer, neurodegeneration, and ischemia-reperfusion injury. BODIPY 581/591 C11 allows researchers to quantify lipid peroxidation in living cells, providing a valuable tool for studying the mechanisms of oxidative stress and ferroptosis, and for screening compounds that modulate these processes. The probe is available from chemical suppliers for research purposes only.

C30H35BF2N2O2

504.4189

504.275

C, 71.43; H, 6.99; B, 2.14; F, 7.53; N, 5.55; O, 6.34

217075-36-0

9914060

Brown to black solid powder

1

5

14

37

934

0

[B-]1(N2C(=CC=C2CCCCCCCCCCC(=O)O)C=C3[N+]1=C(C=C3)/C=C/C=C/C4=CC=CC=C4)(F)F

KXOYQMUYGHQCBT-CMQZKQKJSA-N

InChI=1S/C30H35BF2N2O2/c32-31(33)34-26(17-10-5-3-1-2-4-6-11-19-30(36)37)20-22-28(34)24-29-23-21-27(35(29)31)18-13-12-16-25-14-8-7-9-15-25/h7-9,12-16,18,20-24H,1-6,10-11,17,19H2,(H,36,37)/b16-12+,18-13+

hydrogen 11-(5,5-difluoro-7-((1E,3E)-4-phenylbuta-1,3-dien-1-yl)-5H-5l4,6l4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborinin-3-yl)undecanoate

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 (24.78 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.)