N,N'-Dinitrosopiperazine
Based on 1 Customer Validation
N,N'-Dinitrosopiperazine (1,4-Dinitrosopiperazine; DNP) is a carcinogen with specificity for nasopharyngeal epithelium and facilitates NPC metastasis. N,N'-Dinitrosopiperazine regulates multiple signaling pathways through protein phosphorylation, including LYRIC at serine 568.
For research use only. We do not sell to patients.
- Purity : 99.96%
- CAS No.: 140-79-4
- Formula: C4H8N4O2
- Molecular Weight:144.13
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Description
In Vitro
N,N'-Dinitrosopiperazine (0.5-100 μM; 48 hours) has no inhibitory effects on the labeled 6-10B cells, and LDH activity is not significantly altered by DNP treatment in the 0.5-8 μM concentration range. However, it is cytotoxic from the concentration 10 μM[1].N,N'-Dinitrosopiperazine (2-8 μM; 24 hours) induces 6-10B cell invasion and motility in a dose-dependent manner. At 6 μM, when compares to the control group, DNP increases cell invasion at 421.7% and cell motility is increased by 328.2%[1].N,N'-Dinitrosopiperazine (6 μM; 24 hours) increases the expression of phospho-LYRIC s568 and LYRIC expression in CNE1 cells[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:The labeled 6-10B cells
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Concentration:0.5, 1, 2, 4, 6, 8, 10, 20, 40, 80, or 100 μM
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Incubation Time:48 hours
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Result:Had no inhibitory effects at the concentration 0-8 μM.
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Cell Line:The NPC cell line CNE1
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Concentration:6 μM
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Incubation Time:24 hours
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Result:Increased phospho-LYRIC s568 and LYRIC expression.
In Vivo
Please do not refer to only one article to determine the experimental conditions. It is recommended to determine the optimal experimental conditions (animal strain, age, dosage, frequency and cycle, detection time and indicators, etc.) through preliminary experiments before the formal experiment.
N,N'-Dinitrosopiperazine (injected into the tail veins; 40 mg/kg; 30 days) inhibits cell motility and invasion, and facilitates NPC metastasis in vivo. From a IHC result, Phospho-LYRIC expression is higher in the metastatic tumors of DNP-treated mice than in those of the untreated control mice[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:BABL/c nude mice injected with labeled 6-10B cell suspensions (1 × 104 cells) with or without DNP(40 mg/kg)[1]
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Dosage:40 mg/kg
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Administration:Injected into the tail veins; 30 days
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Result:Induced LYRIC phosphorylation at serine 568 associated with NPC metastasis in vivo.
Chemical Information
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CAS No. 140-79-4
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Appearance Solid
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Molecular Weight 144.13
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Formula C4H8N4O2
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Color White to off-white
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SMILES
O=NN1CCN(N=O)CC1
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Synonyms
1,4-Dinitrosopiperazine; DNP
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Solvent & Solubility
In Vitro:
DMSO : 100 mg/mL (693.82 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (17.35 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (17.35 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocols
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Kinase activity and phosphorylation assays
Kinase activity assays measure the ability of kinases to transfer phosphate groups from ATP to specific substrates, while phosphorylation assays detect the presence and levels of phosphorylated proteins. Common methods include radiolabeled ATP incorporation (e. g. ,), ADP release detection via bioluminescence (e. g. ,[3]), enzyme-linked immunosorbent assays (ELISA) for phospho-specific epitopes (e. g. ,[6]), and microtiter-based formats for high-throughput screening (e. g. ,[8]). The ADP-Glo assay quantifies kinase activity by measuring ADP produced during phosphorylation using a luciferase-based system. Radiometric assays involve autoradiography or scintillation counting after incorporation of 32P-labeled ATP into substrate proteins. ELISA-based approaches rely on phospho-specific antibodies to detect activated kinases in cell lysates or purified samples.
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Western Blot
Western blotting (WB) is a commonly used experimental method in molecular biology, biochemistry, and immunogenetics for identifying and quantifying target proteins. It combines gel electrophoresis with immunoassay, enabling researchers to analyze protein expression, post-translational modifications, and molecular weight.
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Human pluripotent stem cell neural induction and neuron differentiation
Human pluripotent stem cell neural induction can be achieved by blocking BMP and TGFβ/Activin/Nodal SMAD signaling, which suppresses non-neural differentiation and promotes early neuroectodermal identity; the expected readout is loss of pluripotency markers such as OCT4 and induction of neural markers such as PAX6, followed by neural progenitor and neuron marker acquisition during differentiation. This protocol uses dual-SMAD neural induction as the core induction method, followed by cortical neuron differentiation as a representative neuron differentiation model; published cortical protocols describe generation of cortical progenitors, temporally ordered cortical projection neurons, action-potential firing, synaptogenesis, and neural network formation over an approximately 80-day process.
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Protocol for Kinase activity and phosphorylation assays
Kinase activity assays measure transfer of phosphate from ATP to a protein or peptide substrate, generating phosphorylated substrate, ADP, or incorporated radiolabeled phosphate as the readout; phosphorylation assays measure site-specific phosphorylation in cells or tissues as a proxy for kinase-pathway activation, inhibition, or substrate regulation. Phosphorylation can be detected by phospho-specific Western blot, immunoprecipitation kinase assay, phospho-immunofluorescence, phospho-flow cytometry, luminescent ADP detection, radiolabeled ATP incorporation, or reporter-based pathway assays, and these readouts can be applied to cancer cells, primary neurons, mouse tumors, organoids, inflammatory macrophages, ferroptosis studies, and mitophagy studies when the kinase target is biologically relevant.
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Orthotopic Cell-Line Xenograft
Orthotopic cell-line xenograft models involve implantation of human cancer cell lines into the anatomically corresponding organ of immunodeficient mice to reproduce tumor growth within a native microenvironment, enabling more clinically relevant tumor behavior compared with subcutaneous models. These models are widely used because orthotopic placement better recapitulates tumor progression, including invasion and metastatic spread, which are often underrepresented in heterotopic implantation systems. Compared with conventional xenografts, orthotopic implantation is described as more technically complex but provides improved simulation of tumor-microenvironment interactions and metastatic behavior, making it particularly valuable for translational oncology research. Surgical orthotopic implantation approaches have been emphasized as enabling faithful reproduction of clinical cancer features, including metastasis and disease progression patterns that align with the tumor’s organ of origi
Purity & Documentation
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Data Sheet (273 KB)
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SDS (645 KB)
- English - EN (645 KB)
- Français - FR (645 KB)
- Deutsch - DE (645 KB)
- Norwegian - NO (645 KB)
- Español - ES (645 KB)
- Swedish - SV (645 KB)
- Italian - IT (645 KB)
- Korean - KR (645 KB)
- Portuguese - PT (645 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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| DMSO | 1 mM | 6.9382 mL | 34.6909 mL | 69.3818 mL | 173.4545 mL |
| 5 mM | 1.3876 mL | 6.9382 mL | 13.8764 mL | 34.6909 mL | |
| 10 mM | 0.6938 mL | 3.4691 mL | 6.9382 mL | 17.3455 mL | |
| 15 mM | 0.4625 mL | 2.3127 mL | 4.6255 mL | 11.5636 mL | |
| 20 mM | 0.3469 mL | 1.7345 mL | 3.4691 mL | 8.6727 mL | |
| 25 mM | 0.2775 mL | 1.3876 mL | 2.7753 mL | 6.9382 mL | |
| 30 mM | 0.2313 mL | 1.1564 mL | 2.3127 mL | 5.7818 mL | |
| 40 mM | 0.1735 mL | 0.8673 mL | 1.7345 mL | 4.3364 mL | |
| 50 mM | 0.1388 mL | 0.6938 mL | 1.3876 mL | 3.4691 mL | |
| 60 mM | 0.1156 mL | 0.5782 mL | 1.1564 mL | 2.8909 mL | |
| 80 mM | 0.0867 mL | 0.4336 mL | 0.8673 mL | 2.1682 mL | |
| 100 mM | 0.0694 mL | 0.3469 mL | 0.6938 mL | 1.7345 mL |
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