1. Apoptosis
  2. TNF-alpha

Necrostatin 2 

Cat. No.: HY-14622 Purity: 99.89% ee.: 99.20%
Data Sheet SDS Handling Instructions

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.

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Necrostatin 2 Chemical Structure

Necrostatin 2 Chemical Structure

CAS No. : 852391-19-6

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Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO $154 In-stock
5 mg $140 In-stock
10 mg $240 In-stock
50 mg $540 In-stock
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Other Forms of Necrostatin 2:

  • Biological Activity

  • Protocol

  • Technical Information

  • Purity & Documentation

  • References

Description

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.

IC50 & Target

Necroptosis[1]

In Vitro

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].

References
Preparing Stock Solutions
Concentration Volume (DMSO) Mass 1 mg 5 mg 10 mg
1 mM 3.6009 mL 18.0044 mL 36.0088 mL
5 mM 0.7202 mL 3.6009 mL 7.2018 mL
10 mM 0.3601 mL 1.8004 mL 3.6009 mL
Kinase Assay
[2]

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]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Assay
[2]

Necrostatin 2 is dissolved in DMSO and stored, and then diluted with appropriate medium before use[2].

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]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

References
Molecular Weight

277.71

Formula

C₁₃H₁₂ClN₃O₂

CAS No.

852391-19-6

Storage
Powder -20°C 3 years
  4°C 2 years
In solvent -80°C 6 months
  -20°C 1 month
Shipping

Room temperature in continental US; may vary elsewhere

Solvent & Solubility

10 mM in DMSO

* "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

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Necrostatin 2
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HY-14622
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