| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
The target is intracellular hydrogen peroxide (H2O2), a key reactive oxygen species (ROS). The probe's mechanism is based on a specific chemical reaction: H2O2 cleaves the boronic ester groups, triggering a self-immolative process that releases the highly fluorescent fluorescein. This design ensures that the signal is specific to H2O2 and not other ROS.
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| ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular requirements as it simply offers guidance) [1]. 1. In a 35 mm culture dish with a glass bottom, seed RAW 264.7 cells at a density of 250,000 cells and let them post-adhere for 5 hours. 2. Apply 1 μg/mL LPS to a few of the culture dishes for a whole day. The day following plating, 100 μM H2O2 or PBS were applied to the remaining dishes for one hour each. 3. After giving cells two PBS washes, fix them for fifteen minutes in ice-cold 95% ethanol. 3. After giving cells two PBS washes, incubate them for an hour at room temperature with a 50 μM probe (FBBBE). 4. After giving the cells two PBS washes, use Aqua Mount to attach the coverslip to the cells. 5. Using an Olympus FV1000 confocal laser scanning microscope with an 80x magnification, confocal pictures were captured. 6. To process the image, use the ImageJ program.
FBBBE is a probe, not a drug. Its "activity" is the increase in fluorescence upon reaction with H2O2, which is measured as a proxy for cellular ROS levels. In solution, it shows a significant turn-on fluorescence response. The compound is virtually non-fluorescent until it is cleaved by H2O2. It can specifically detect intracellular, but not extracellular, H2O2, due to its cell-permeability. |
| ln Vivo |
Not applicable. This is a diagnostic imaging probe and does not exert a therapeutic biological effect in vivo. Its "in vivo activity" is its ability to generate a fluorescent signal upon encountering H2O2, but its use is primarily for in vitro cell-based assays due to rapid clearance and autofluorescence issues in whole animals.
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| Enzyme Assay |
Not applicable. As a chemical probe that reacts with a small molecule (H2O2), it is not evaluated in a classical enzyme/receptor binding assay. Its performance is evaluated in a cuvette using buffer solutions. A standard "activity" test involves adding H2O2 to a solution of FBBBE and measuring the increase in fluorescence (Ex/Em ~480/512 nm) over time using a fluorometer.
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| Cell Assay |
The standard protocol for cellular assays: 1. Seed cells (e.g., HeLa, macrophages) in a 96-well plate or on a coverslip. 2. Remove culture medium and wash cells with PBS. 3. Load cells with FBBBE (1-10 microM) in serum-free medium for 30-60 minutes at 37degC in the dark. 4. Remove the loading buffer and wash cells to remove excess probe. 5. Stimulate ROS production by adding an agonist (e.g., PMA at 100 nM, or H2O2 as a positive control). 6. Measure fluorescence intensity immediately or over time using a fluorescence plate reader (Ex/Em=480/512 nm) or capture images via confocal microscopy.
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| Animal Protocol |
Not applicable. FBBBE is designed specifically for in vitro assays. It is not used in animal models due to the high background fluorescence and rapid metabolism of fluorescein derivatives in vivo. Typical in vivo ROS probes require different chemistries for biodistribution and stability.
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| ADME/Pharmacokinetics |
Not applicable. This is an imaging probe for in vitro use. Pharmacokinetic properties are not relevant as it is not intended for systemic delivery. The key property is its cell permeability, which allows it to cross the intact plasma membrane of living cells to detect intracellular H2O2.
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| Toxicity/Toxicokinetics |
Not applicable (non-toxic fluorescent probe at working concentrations). Standard handling and disposal protocols for organic solvents (DMSO) should be followed. The compound itself is used as a diagnostic and does not induce cytotoxicity in cells during the short-term loading and imaging periods described in standard protocols.
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| References | |
| Additional Infomation |
Not applicable. FBBBE is a research tool for studying oxidative stress and redox biology in cell culture. It is a "turn-on" probe that allows for real-time monitoring of intracellular H2O2 production. It is not a drug candidate and therefore has no clinical trials or approval status. It is a chemical reagent for laboratory use.
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| Molecular Formula |
C46H46B2O9
|
|---|---|
| Molecular Weight |
764.474
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| Exact Mass |
584.238
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| CAS # |
1522117-83-4
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| PubChem CID |
77461126
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| Appearance |
Off-white to light yellow solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
595.2±60.0 °C at 760 mmHg
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| Flash Point |
313.8±32.9 °C
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| Vapour Pressure |
0.0±1.7 mmHg at 25°C
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| Index of Refraction |
1.600
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
57
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| Complexity |
1320
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O=C1C2=CC=CC=C2C2(C3=CC=C(OCC4C=CC(B5OC(C)(C)C(C)(C)O5)=CC=4)C=C3OC3=CC(OCC4C=CC(B5OC(C)(C)C(C)(C)O5)=CC=4)=CC=C23)O1
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| InChi Key |
UKXLQKOGCSYSOD-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C46H46B2O9/c1-42(2)43(3,4)55-47(54-42)31-17-13-29(14-18-31)27-50-33-21-23-37-39(25-33)52-40-26-34(22-24-38(40)46(37)36-12-10-9-11-35(36)41(49)53-46)51-28-30-15-19-32(20-16-30)48-56-44(5,6)45(7,8)57-48/h9-26H,27-28H2,1-8H3
|
| Chemical Name |
3',6'-bis[[4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]methoxy]spiro[2-benzofuran-3,9'-xanthene]-1-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 |
| 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 | 1.3081 mL | 6.5405 mL | 13.0810 mL | |
| 5 mM | 0.2616 mL | 1.3081 mL | 2.6162 mL | |
| 10 mM | 0.1308 mL | 0.6540 mL | 1.3081 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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