Necrostatin 2
Based on 11 publication(s) in Google Scholar
Necrostatin 2 is a potent necroptosis inhibitor. EC50 for inhibition of necroptosis in FADD-deficient Jurkat T cells treated with TNF-α is 0.05 μM. Necrostatin 2 is also a RIPK1 inhibitor.
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- Pureté: 99.78%
- CAS No.: 852391-19-6
- Formule: C13H12ClN3O2
- Masse moléculaire:277.71
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Stockage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 2 years , -20°C, 1 year
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Activité biologique
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Chemical Information
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Solvant et solubilité
- Protocole
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Publications Citing Use of MedChemExpress (MCE) Necrostatin 2
More- Mater Today Bio. 2025 Nov 13:35:102547. [Abstract]
- Cell Death Discov. 2026 Feb 23;12(1):105. [Abstract]
- Cell Rep. 2025 Sep 1;44(9):116139. [Abstract]
- Cancer Cell Int. 2024 Jun 3;24(1):194. [Abstract]
- Int Immunopharmacol. 2024 Jun 15:134:112159. [Abstract]
- Mol Carcinog. 2023 Aug;62(8):1119-1135. [Abstract]
- J Cancer. 2023 May 15;14(8):1336-1349. [Abstract]
- FEBS Open Bio. 2019 Feb 23;9(4):582-593. [Abstract]
- Transl Cancer Res. 2021 Dec;10(12):5307-5318. [Abstract]
- Res Sq. 2025 Feb 16.
- St. Johns University. 2021 Jul.
Activité biologique
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RIPK1 |
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Cell Line
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Type | Value | Description | References |
|---|---|---|---|---|
| Jurkat | EC50 |
0.05 μM
Compound: 25
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Effective concentration required for inhibition of necroptosis in FADD deficient Jurkat T cells treated with TNF-alpha
Effective concentration required for inhibition of necroptosis in FADD deficient Jurkat T cells treated with TNF-alpha
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[PMID: 16153840] |
| Jurkat | EC50 |
0.21 μM
Compound: (+/-)-1
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Inhibition of TNF alpha-induced necroptosis in FADD-deficient variant human Jurkat T cells assessed as cell viability after 24 hrs
Inhibition of TNF alpha-induced necroptosis in FADD-deficient variant human Jurkat T cells assessed as cell viability after 24 hrs
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[PMID: 17361994] |
| Jurkat | EC50 |
18 nM
Compound: 11
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Inhibition of endogenous RIP1 autophosphorylation in human Jurkat cells
Inhibition of endogenous RIP1 autophosphorylation in human Jurkat cells
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[PMID: 18408713] |
| Jurkat | EC50 |
50 nM
Compound: 11
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Inhibition of cellular necroptosis in TNFalpha treated FADD deficient human Jurkat cells
Inhibition of cellular necroptosis in TNFalpha treated FADD deficient human Jurkat cells
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[PMID: 18408713] |
| U-937 | IC50 |
320 nM
Compound: 1
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Inhibition of human RIP1 in human U937 cells assessed as inhibition of TNF/zVAD.fmk induced necroptosis after 24 hrs by Cell titer-Glo luminescence assay
Inhibition of human RIP1 in human U937 cells assessed as inhibition of TNF/zVAD.fmk induced necroptosis after 24 hrs by Cell titer-Glo luminescence assay
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[PMID: 26854747] |
Evaluation of necroptosis inhibitory activity is performed using a FADD-deficient variant of human Jurkat T cells treated with TNF-α. Utilizing these conditions the cells efficiently undergo necroptosis, which is completely and selectively inhibited by Necrostatin 2 (EC50=50 nM). Necrostatin 2 shows activity in a broad range of necroptosis cellular systems[1]. Necrostatin 2 at 30 μM completely protects L929 cells from TNF-α-induced necroptosis. In addition to TNF-α, the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp(OMe)-fluoromethylketone (zVAD.fmk) has also been found to induce necrosis in L929 cells, which is efficiently inhibited by Necrostatin 2[2]. EC50 for inhibition of necroptosis in FADD-deficient Jurkat T cells treated with TNF-α is 0.05 μM[3].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 852391-19-6
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Appearance Solid
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Masse moléculaire 277.71
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Formule C13H12ClN3O2
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Color White to light yellow
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SMILES
ClC1=C2C(C(C[C@@H]3C(N(C)C(N3)=O)=O)=CN2)=CC=C1
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Livraison
Room temperature in continental US; may vary elsewhere.
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Stockage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 2 years -20°C 1 year
Publications (11)
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Journal Impact Factor
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Most Recent
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Mater Today Bio
SLC7A11-independent disulfidptosis induced by a two-pronged delivery strategy for bladder cancer chemotherapy and cisplatin-resistance reverse. [Abstract]2025 Nov 13:35:102547. PMID: 41322137 -
Cell Death Discov
PDHA1 enhances resistance to ferroptosis in anoikis-resistant prostate cancer by upregulating AIFM2. [Abstract]2026 Feb 23;12(1):105. PMID: 41724793 -
Cell Rep
SLC6A8-mediated creatine uptake suppresses ERK2-FSP1 signaling and induces ferroptosis in colorectal cancer. [Abstract]2025 Sep 1;44(9):116139. PMID: 40892544 -
Cancer Cell Int
Identifying disulfidptosis subtypes in hepatocellular carcinoma through machine learning and preliminary exploration of its connection with immunotherapy. [Abstract]2024 Jun 3;24(1):194. PMID: 38831301 -
Int Immunopharmacol
Disulfiram mediated anti-tumour effect in pituitary neuroendocrine tumours by inducing cuproptosis. [Abstract]2024 Jun 15:134:112159. PMID: 38692018 -
Mol Carcinog
PRMT1 inhibition promotes ferroptosis sensitivity via ACSL1 upregulation in acute myeloid leukemia. [Abstract]2023 Aug;62(8):1119-1135. PMID: 37144835 -
J Cancer
PERK/ATF3-Reduced ER Stress on high potassium environment in the suppression of tumor ferroptosis. [Abstract]2023 May 15;14(8):1336-1349. PMID: 37283787 -
FEBS Open Bio
2019 Feb 23;9(4):582-593. PMID: 30984534 -
Transl Cancer Res
2021 Dec;10(12):5307-5318. PMID: 35116379 -
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Solvant et solubilité
DMSO : ≥ 100 mg/mL (360.09 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
* "≥" means soluble, but saturation unknown.
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, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
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, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
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 (9.00 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 (9.00 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.
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.
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.
Protocole
Evaluation of necroptosis inhibitory activity is performed using an FADD-deficient variant of human Jurkat T cells treated for 24 h with TNF-α. Under these conditions the cells underwent necroptosis (the DMSO control had ~40% viability), which is inhibited by Necrostatin 2 (EC50=0.21±0.2 μM) as a positive control. For EC50 value determinations, cells are treated with 10 ng/mL human TNF-α in the presence of increasing concentrations of test compounds (0.029, 0.058, 0.12, 0.23, 0.46, 0.93, 1.9, 3.7, 11.1, 33, and 100 μM) in duplicate for 24 h followed by ATP-based viability assessment. EC50 values±SD are determined from at least two independent experiments[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
L929 cells (100000 cells/mL, 100 μL/well in a 96-well plate) are treated with 10 ng/mL human TNF-α or 100 μM zVAD.fmk in the presence of DMSO (control), 30 μM Necrostatin 2, or 8for 24 h at 37°C in a humidified incubator with 5% CO2 followed by ATP-based viability assessment as described in the previous experiment. Stock solutions (30 mM) in DMSO are initially prepared and then diluted with DMSO to give testing solutions. Each sample is done in duplicate. The final DMSO concentration is 0.5%. Cell viability values are adjusted to account for nonspecific toxicity, which in most cases is <10%. The reported cell viability values (%)±SD are determined from two independent experiments[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Pureté et documentation
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Fiche technique (283 KB)
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SDS (394 KB)
- English - EN (394 KB)
- Français - FR (394 KB)
- Deutsch - DE (394 KB)
- Norwegian - NO (394 KB)
- Español - ES (394 KB)
- Swedish - SV (394 KB)
- Italian - IT (394 KB)
- Korean - KR (394 KB)
- Portuguese - PT (394 KB)
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Instruction de manipulation (2659 KB)
Références
[1]. Teng X, et al. Structure-activity relationship study of [1,2,3]thiadiazole necroptosis inhibitors. Bioorg Med Chem Lett. 2007 Dec 15;17(24):6836-40. [Content Brief]
[2]. Jagtap PG, et al. Structure-activity relationship study of tricyclic necroptosis inhibitors. J Med Chem. 2007 Apr 19;50(8):1886-95. [Content Brief]
[3]. Teng X, et al. Structure-activity relationship study of novel necroptosis inhibitors. Bioorg Med Chem Lett. 2005 Nov 15;15(22):5039-44. [Content Brief]
[4]. Takahashi N, et al. Necrostatin-1 analogues: critical issues on the specificity, activity and in vivo use in experimental disease models. Cell Death Dis. 2012 Nov 29;3:e437. [Content Brief]
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, 2 years; -20°C, 1 year. When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 3.6009 mL | 18.0043 mL | 36.0087 mL | 90.0216 mL |
| 5 mM | 0.7202 mL | 3.6009 mL | 7.2017 mL | 18.0043 mL | |
| 10 mM | 0.3601 mL | 1.8004 mL | 3.6009 mL | 9.0022 mL | |
| 15 mM | 0.2401 mL | 1.2003 mL | 2.4006 mL | 6.0014 mL | |
| 20 mM | 0.1800 mL | 0.9002 mL | 1.8004 mL | 4.5011 mL | |
| 25 mM | 0.1440 mL | 0.7202 mL | 1.4403 mL | 3.6009 mL | |
| 30 mM | 0.1200 mL | 0.6001 mL | 1.2003 mL | 3.0007 mL | |
| 40 mM | 0.0900 mL | 0.4501 mL | 0.9002 mL | 2.2505 mL | |
| 50 mM | 0.0720 mL | 0.3601 mL | 0.7202 mL | 1.8004 mL | |
| 60 mM | 0.0600 mL | 0.3001 mL | 0.6001 mL | 1.5004 mL | |
| 80 mM | 0.0450 mL | 0.2251 mL | 0.4501 mL | 1.1253 mL | |
| 100 mM | 0.0360 mL | 0.1800 mL | 0.3601 mL | 0.9002 mL |
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