Search Site
Home >> Signaling Pathways >> Apoptosis >> TNF-α >> Necrostatin-1
Related Products
Necrostatin-1RIP1 inhibitor

Necrostatin-1

Catalog No. A4213
Size Price Stock Qty
10mM (in 1mL DMSO) $55.00 In stock
10mg $45.00 In stock
100mg $190.00 In stock

Tel: +1-832-696-8203

Email: sales@apexbt.com

Worldwide Distributors

Sample solution is provided at 25 µL, 10mM.

Quality Control

Chemical structure

Necrostatin-1

Related Biological Data

Necrostatin-1

Biological Activity

Description Necrostatin-1 is a specific inhibitor of RIP1 and inhibits TNF-α-induced necroptosis with an EC50 value of 490 nM.
Targets RIP1          
IC50 490 nM (EC50)          

Protocol

Kinase experiment [1]:

RIP1 kinase assay

pcDNA3-FLAG-RIP1 vector is transfected into 293T cells and RIP1 kinase assay is performed in the presence of 10 μM cold ATP and 1 μCi of [γ-32P]ATP for 30 min at 30℃. Reactions were stopped by boiling in SDS-PAGE sample buffer. Samples are subjected to SDS-PAGE and RIP1 band is visualized by autoradiography.

Cell experiment [2]:

Cell lines

Mouse osteocyte cell line (MLO-Y4)

Preparation method

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reacting condition

30 mM; 24h

Applications

Necrostatin-1 (30 mmol/L) inhibited the mouse osteocyte cell line (MLO-Y4) necroptosis induced by TNF-α in vitro.

Animal experiment [2, 3]:

Animal models

Rats underwent the ovariectomized surgery; Eight-week-old mice underwent sham surgery or contrast-induced AKI treatment;

Dosage form

1.65 mg/kg/d; intraperitoneal injection; once per day for 4 weeks

Applications

Treatment with Necrostatin-1 (1.65 mg/kg/d) significantly decreased RIP1 and RIP3 expression in ovariectomized rats. Moreover, necrostatin-1 prevented osmotic nephrosis and contrast-induced AKI in mice.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

1Degterev, A., Hitomi, J., Germscheid, M., Ch'en, I. L., Korkina, O., Teng, X., Abbott, D., Cuny, G. D., Yuan, C., Wagner, G., Hedrick, S. M., Gerber, S. A., Lugovskoy, A. and Yuan, J. (2008) Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol. 4, 313-3212Cui, H., Zhu, Y., Yang, Q., Zhao, W., Zhang, S., Zhou, A. and Jiang, D. (2016) Necrostatin-1 treatment inhibits osteocyte necroptosis and trabecular deterioration in ovariectomized rats. Sci Rep. 6, 338033Linkermann, A., Heller, J. O., Prokai, A., Weinberg, J. M., De Zen, F., Himmerkus, N., Szabo, A. J., Brasen, J. H., Kunzendorf, U. and Krautwald, S. (2013) The RIP1-kinase inhibitor necrostatin-1 prevents osmotic nephrosis and contrast-induced AKI in mice. J Am Soc Nephrol. 24, 1545-1557

2Cui, H., Zhu, Y., Yang, Q., Zhao, W., Zhang, S., Zhou, A. and Jiang, D. (2016) Necrostatin-1 treatment inhibits osteocyte necroptosis and trabecular deterioration in ovariectomized rats. Sci Rep. 6, 33803

3Linkermann, A., Heller, J. O., Prokai, A., Weinberg, J. M., De Zen, F., Himmerkus, N., Szabo, A. J., Brasen, J. H., Kunzendorf, U. and Krautwald, S. (2013) The RIP1-kinase inhibitor necrostatin-1 prevents osmotic nephrosis and contrast-induced AKI in mice. J Am Soc Nephrol. 24, 1545-1557

Necrostatin-1 Dilution Calculator

Concentration (start)
x
Volume (start)
=
Concentration (final)
x
Volume (final)
 
 
 
C1
V1
C2
V2

calculate

Necrostatin-1 Molarity Calculator

Mass
=
Concentration
x
Volume
x
MW*
 
 
 
g/mol

calculate

Chemical Properties

Cas No. 4311-88-0 SDF Download SDF
Synonyms MTH-DL-Tryptophan,Nec-1
Chemical Name 5-(1H-indol-3-ylmethyl)-3-methyl-2-sulfanylideneimidazolidin-4-one
Canonical SMILES CN1C(=O)C(NC1=S)CC2=CNC3=CC=CC=C32
Formula C13H13N3OS M.Wt 259.33
Solubility >13mg/mL in DMSO Storage Store at -20°C
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

Background

IC50: Necrostatin-1 (Nec-1), (R)-5-([7-chloro-1H-indol-3-yl]methyl)-3-methylimidazolidine-2,4-dione (Nec-1a) (Figure 1A) (Degterev et al., 2008), exhibited an inhibitory constant (IC50) of 0.32 mM for RIP1 [1].

Necroptosis is a cellular mechanism of necrotic cell death induced by apoptotic stimuli in the form of death domain receptor engagement by their respective ligands under conditions where apoptotic execution is prevented. Necrostatin-1, identified as a small-molecule inhibitor of necroptosis, is also a selective allosteric inhibitor of the death domain receptor–associated adaptor kinase RIP1.

In vitro: Previous study indicated that necrostatin-1 was a selective allosteric inhibitor of the death domain receptor–associated adaptor kinase RIP1 in vitro. In this study, RIP1 was found to be the primary cellular target responsible for the antinecroptosis activity of necrostatin-1. In addition, two other necrostatins, necrostatin-3 and necrostatin-5, were also shown to target the RIP1 kinase step in the necroptosis pathway, but through different mechanism compared with that of necrostatin-1. The findings established necrostatins as the first-in-class inhibitors of RIP1 kinase, the key upstream kinase involved in the activation of necroptosis [2].

In vivo: A previous study was designed to investigate the protective effects and mechanisms of Nec-1 in concanavalin A-induced hepatitis in mice. It was found that in Nec-1-treated mice the amelioration in liver functions and histopathological changes and the suppression of inflammatory cytokine production were observed. Western blotting analyses showed that the expression of TNF-α, IFN-γ, IL2, IL6, and RIP1 was significantly reduced in the Nec-1-treated mice, which was further confirmed by immunofluorescence and immunohistochemistry. In addition, autophagosome formation was significantly reduced by Nec-1 treatment. These results indicated that Nec-1 could prevent concanavalin A -induced liver injury via RIP1-related and autophagy-related pathways [3].

Clinical trial: Up to now, Necroptosis is still in the preclinical development stage.

References:
[1] Xie T, Peng W, Liu Y, Yan C, Maki J, Degterev A, Yuan J, Shi Y.  Structural basis of RIP1 inhibition by necrostatins. Structure. 2013;21(3):493-9.
[2] Degterev A, Hitomi J, Germscheid M, Ch'en IL, Korkina O, Teng X, Abbott D, Cuny GD, Yuan C, Wagner G, Hedrick SM, Gerber SA, Lugovskoy A, Yuan J.  Identification of RIP1 kinase as a specific cellular target of necrostatins. Nat Chem Biol. 2008;4(5):313-21.
[3] Yingqun Zhou, Weiqi Dai, Chunlei Lin, Fan Wang, Lei He, Miao Shen, Ping Chen, Chenfen Wang, Jie Lu, Ling Xu, Xuanfu Xu, and Chuanyong Guo.  Protective Effects of Necrostatin-1 against Concanavalin A-Induced Acute Hepatic Injury in Mice. Mediators of Inflammation. http://dx.doi.org/10.1155/2013/706156