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| Targets |
NucPE1 targets hydrogen peroxide (H2O2) in the nucleus of living cells. The probe is specifically localized to cellular nuclei without appended targeting moieties. The boronate-based probe reacts selectively with hydrogen peroxide, resulting in a fluorescent product. The compound's nuclear localization enables the specific detection of nuclear H2O2 levels, which is important for studying oxidative stress, DNA damage, and redox signaling in the nucleus. NucPE1's enhanced emission properties (λem=530 nm, Φ=0.626) and major absorption band at 505 nm (ε=19,100 M-¹cm-¹) make it suitable for fluorescence microscopy and flow cytometry applications.
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| ln Vitro |
NucPE1 exhibits weak emission (λem=530 nm, Φ=0.117) and two main visible region absorptions (λabs=468 nm, ε=27,300 M-1cm-1; λabs=490 nm, ε=26,000 M-1cm-1). When NucPE1 reacts with H2O2, it forms the fluorophore NucPE1, which has enhanced emission (λem=530 nm, Φ =0.626) and a major absorption band at 505 nm (ε = 19,100 M-1cm-1). This reaction causes fluorescence enhancement. NucPE1 preferentially localizes in the nuclei of various mammalian cell lines and entire model organisms, including C. It can react to subcellular variations in H2O2 flux in clegans [1].
NucPE1 demonstrates in vitro activity as a fluorescent hydrogen peroxide probe. The compound is a nuclear-localized fluorescent probe that specifically detects H2O2 in the nucleus. NucPE1 features enhanced emission (λem=530 nm, Φ=0.626) and a major absorption band at 505 nm (ε=19,100 M-¹cm-¹). The probe reacts selectively with hydrogen peroxide, producing a fluorescent signal that can be detected by fluorescence microscopy or flow cytometry. NucPE1 imaging reveals a reduction in nuclear H2O2 levels in response to various stimuli. The compound's activity is concentration-dependent, with optimal staining observed at appropriate concentrations. |
| ln Vivo |
In vivo, NucPE1 is still able to target the nucleus specifically. Nuclear H2O2 levels were lower in C, according to NucPE1 imaging. elegans overexpressing Sir-2.1 in comparison to the homolog of the wild type, indicating a connection between improved control of nuclear ROS pools and this long-lived Sirtuin protein [1].
NucPE1 is used for in vivo imaging of nuclear hydrogen peroxide in living cells and organisms. The probe specifically localizes to cellular nuclei without appended targeting moieties. NucPE1 imaging reveals a reduction in nuclear H2O2 levels in response to various stimuli. The compound's nuclear localization and fluorescent properties make it suitable for studying oxidative stress, DNA damage, and redox signaling in the nucleus of living cells and tissues. NucPE1 can be used in various biological models to investigate the role of nuclear H2O2 in physiological and pathological processes. |
| Enzyme Assay |
In vitro assays for NucPE1 involve measuring its fluorescent response to hydrogen peroxide. The compound is incubated with varying concentrations of H2O2 in buffer solutions, and fluorescence is measured using a fluorescence plate reader with excitation at 505 nm and emission at 530 nm. The fluorescence enhancement is plotted against H2O2 concentration to determine the probe's sensitivity and dynamic range. Selectivity assays compare the compound's response to H2O2 versus other reactive oxygen species (ROS) and reactive nitrogen species (RNS). The compound's structure and purity can be characterized using analytical methods. High-performance liquid chromatography (HPLC) and mass spectrometry are used to verify the molecular weight (483.36 g/mol) and chemical composition (C29H30BNO5). Purity (≥98%) is confirmed by HPLC analysis.
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| Cell Assay |
In vitro cellular assays for NucPE1 are performed using various cell lines to detect nuclear hydrogen peroxide. Cells are loaded with NucPE1 by incubation with the probe for a defined period. After washing, cells are treated with stimuli that induce H2O2 production (e.g., growth factors, inflammatory cytokines, or oxidative stress inducers). Nuclear fluorescence is visualized using fluorescence microscopy with excitation at 514 nm and emission collected between 522-554 nm. Fluorescence intensity is quantified using image analysis software or flow cytometry. Cell viability is assessed in parallel using standard viability assays to ensure that probe loading does not affect cell viability. The probe's nuclear localization is confirmed by co-staining with nuclear dyes such as Hoechst or DAPI.
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| Animal Protocol |
In vivo animal studies for NucPE1 are conducted to image nuclear hydrogen peroxide in living organisms. The probe is administered via intravenous injection or local application, and fluorescence imaging is performed using appropriate excitation and emission filters. NucPE1 imaging reveals nuclear H2O2 levels in various tissues. The compound's nuclear localization and fluorescent properties make it suitable for studying oxidative stress and redox signaling in vivo. Pharmacokinetic studies assess probe distribution and clearance. Animals are monitored for clinical signs and body weight. The probe's utility for in vivo imaging has been demonstrated in research publications.
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| ADME/Pharmacokinetics |
Pharmacokinetic properties of NucPE1 have been characterized in research studies. The compound has a molecular formula of C29H30BNO5 and a molecular weight of 483.36 g/mol. Its chemical name is 3'-(ethylamino)-2'-methyl-6'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one. NucPE1 is soluble in DMSO and other organic solvents. The compound should be stored at 2-8degC, protected from light, and under argon. Comprehensive pharmacokinetic parameters including half-life, volume of distribution, clearance, and bioavailability have been characterized in preclinical studies. The compound's stability and fluorescent properties support its use as a research probe.
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| Toxicity/Toxicokinetics |
NucPE1 is intended for laboratory research use only and has not undergone comprehensive toxicology testing. As a fluorescent probe, the compound is used in small quantities for imaging applications. Standard in vitro cytotoxicity assays in cell lines are typically performed alongside imaging studies to ensure that probe loading does not affect cell viability. In vivo, animals are monitored for signs of toxicity including body weight changes and clinical observations. Comprehensive toxicological characterization including genotoxicity and repeated-dose toxicity studies has not been reported. The probe is not approved for human use and is strictly intended for research purposes.
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| References |
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| Additional Infomation |
NucPE1 (Nuclear Peroxy Emerald 1) is a nuclear-localized fluorescent hydrogen peroxide probe. It specifically localizes to the nucleus without an additional targeting moiety. NucPE1 has enhanced emission (λem=530 nm, Φ=0.626) and a major absorption band at 505 nm (ε=19,100 M-¹cm-¹). The compound has a molecular formula of C29H30BNO5 and a molecular weight of 483.36 g/mol. NucPE1 imaging reveals a reduction in nuclear H2O2 levels. The compound has not entered clinical trials and is available from research chemical suppliers for non-clinical research purposes only.
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| Molecular Formula |
C29H30BNO5
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| Molecular Weight |
483.3632
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| Exact Mass |
483.221
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| CAS # |
1404091-23-1
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| PubChem CID |
129626434
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
36
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| Complexity |
849
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O1B(C2C([H])=C([H])C3=C(C=2[H])OC2C([H])=C(C(C([H])([H])[H])=C([H])C=2C23C3=C([H])C([H])=C([H])C([H])=C3C(=O)O2)N([H])C([H])([H])C([H])([H])[H])OC(C([H])([H])[H])(C([H])([H])[H])C1(C([H])([H])[H])C([H])([H])[H]
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| InChi Key |
GHPDUBPWQBBRPG-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C29H30BNO5/c1-7-31-23-16-25-22(14-17(23)2)29(20-11-9-8-10-19(20)26(32)34-29)21-13-12-18(15-24(21)33-25)30-35-27(3,4)28(5,6)36-30/h8-16,31H,7H2,1-6H3
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| Chemical Name |
3'-(ethylamino)-2'-methyl-6'-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture. |
| 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) |
DMSO : ~25 mg/mL (~51.72 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.17 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0689 mL | 10.3443 mL | 20.6885 mL | |
| 5 mM | 0.4138 mL | 2.0689 mL | 4.1377 mL | |
| 10 mM | 0.2069 mL | 1.0344 mL | 2.0689 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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