Necrostatin-1

Catalog No.S8037 Synonyms: Nec-1

For research use only.

Necrostatin-1 (Nec-1) is a specific RIP1 (RIPK1) inhibitor and inhibits TNF-α-induced necroptosis with EC50 of 490 nM in 293T cells. Necrostatin-1 also blocks IDO and suppresses autophagy and apoptosis.

Necrostatin-1 Chemical Structure

CAS No. 4311-88-0

Selleck's Necrostatin-1 has been cited by 126 publications

Purity & Quality Control

Choose Selective RIP kinase Inhibitors

Biological Activity

Description Necrostatin-1 (Nec-1) is a specific RIP1 (RIPK1) inhibitor and inhibits TNF-α-induced necroptosis with EC50 of 490 nM in 293T cells. Necrostatin-1 also blocks IDO and suppresses autophagy and apoptosis.
Features A powerful tool for characterizing the role of necroptosis with characterized primary target.
Targets
RIP1 [1]
(293T cells)
490 nM(EC50)
In vitro

Necrostatin-1 (1-100 μM) inhibits the autophosphorylation of overexpressed and endogenous RIP1.It is found RIP1 is the primary cellular target responsible for the antinecroptosis activity of Necrostatin-1. [1] Necrostatin-1 efficiently suppresses necroptotic cell death triggered by an array of stimuli in a variety of cell types. Necrostatin-1, previously identified as small-molecule inhibitor of necroptosis, inhibits RIP kinase-induced necroptosis and inhibits TNF-α-induced necroptosis in jurkat cells with EC50 of 490 nM. [2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
HT-22 NVHvcYdPS3m2b4TvfIlkcXS7IFHzd4F6 M1vmSlExKM7:TR?= NWLEe49LOTJiaB?= NUT5OmFOeHKxdHXjeJMh[WejaX7zeEBk\WyuIHTlZZRpKGmwZIXj[YQh[nliNdMgcY1wdC:OIHfseZRidWG2ZdMg M1H3W|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF5N{[wPFY6Lz5zN{e2NFg3QTxxYU6=
Jurkat  Mn\qSpVv[3Srb36gRZN{[Xl? MX:yNFDDqM7:bR?= MVizNEBucW5? MUjy[YR2[2W|wrDOZYVodGW{aXGg[o94dGW{aT3pcoR2[2WmIILlZYN1cX[nIH;4fYdmdiC|cHXjbYV{KCiUT2OpJIdmdmW{YYTpc44> M17QVFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzNUO1NFIxLz5{MUWzOVAzODxxYU6=
Jurkat  MoraR5l1d3SxeHnjbZR6KEG|c3H5 NWK4T|lVPTBxIEGwNE8zODEEoN88cS=> NGHNTW4yNzNiaB?= NHTCdpVz\WS3Y3XzxsBP[WWpbHXybYEh\m:5bHXybU1qdmS3Y3XkJIN6fG:2b4jpZ4l1gQ>? MmH3QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjF3M{WwNlAoRjJzNUO1NFIxRC:jPh?=
SW13 M4PVW2NmdGxiVnnhZoltcXS7IFHzd4F6 NYX5Sm9kOTByIN88US=> NInPNYszPCCq M4f2dWROW09? Mn3VbY5kemWjc3XzJINmdGy3bHHyJJN2en[rdnHs M4TCbFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{MUO2PFE5Lz5{MkGzOlgyQDxxYU6=
8505c NH;vO2JE\WyuIG\pZYJqdGm2eTDBd5NigQ>? M2jlTFExOCEQvF2= NGPSUm4zPCCq MV7EUXNQ M3TmZ4lv[3KnYYPld{Bk\WyudXzhdkB{fXK4aY\hcC=> M4GyWVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{MUO2PFE5Lz5{MkGzOlgyQDxxYU6=
TPC-1 MYnD[YxtKF[rYXLpcIl1gSCDc4PhfS=> NVPn[4pxOTByIN88US=> MWOyOEBp M1;OVGROW09? MkLhbY5kemWjc3XzJINmdGy3bHHyJJN2en[rdnHs NFfPb4s9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkGzOlgyQCd-MkKxN|Y5OTh:L3G+
L929sA MVfBdI9xfG:|aYOgRZN{[Xl? NIn2RYsyOCEQvF2= M{PFPFEhcA>? MlXYZYJzd2ejdHXzJJRp\SCrboTldoFkfGmxbjDv[kBk[XOyYYPlMVghf2m2aDDGRWRF NX;QbG5HRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkKzOlI4PjdpPkKyN|YzPzZ5PD;hQi=>
L929sA M3P5SGFxd3C2b4Ppd{BCe3OjeR?= NHPqc40yOCEQvF2= NEf0XJYyKGh? NYCzWlJZemW|Y4Xld{Bk\WyuczDlfJBz\XO|aX7nJHJKWEtzzqTJSEBnem:vIGTOSk1qdmS3Y3XkJIFxd3C2b4Ppdy=> MWe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjN4Mke2O{c,OjJ|NkK3Olc9N2F-
L929sA NX3kNmJ5SXCxcITvd4l{KEG|c3H5 NWrVZYF7OTBizszN NF;tVJIyKGh? MUTpcohq[mm2czD0bIUh[XCxcITveIlkKHKnc4DvcpNmKHSxIGTOSi=> NIm1SHY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkO2Nlc3Pyd-MkKzOlI4Pjd:L3G+
K562/Adr  M3zNZWZ2dmO2aX;uJGF{e2G7 NYnGcI9YPjBizszN M4HZPVEzKGh? MmX2bY5kemWjc3XzJJRp\SCjY4Tpeol1gSCxZjDjZZNx[XOnczygZ4F{eGG|ZTC4JIFv\CB7 NYXiTYZyRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK4N|c3QDlpPkKyPFM4Pjh7PD;hQi=>
K562 MVXGeY5kfGmxbjDBd5NigQ>? MlTIOlAh|ryP NF7F[moyOiCq NHXzXYFqdmO{ZXHz[ZMhfGinIHHjeIl3cXS7IH;mJINie3Cjc3XzMEBk[XOyYYPlJFgh[W6mIEm= NYnJeW92RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK4N|c3QDlpPkKyPFM4Pjh7PD;hQi=>
HL60/Adr NFXmeHhHfW6ldHnvckBCe3OjeR?= M{Xv[lYxKM7:TR?= MmD6NVIhcA>? NIXMe5VqdmO{ZXHz[ZMhfGinIHHjeIl3cXS7IH;mJINie3Cjc3XzMEBk[XOyYYPlJFgh[W6mIEm= M2nLWlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{OEO3Olg6Lz5{MkizO|Y5QTxxYU6=
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K562/Adr  M4XvRWZ2dmO2aX;uJGF{e2G7 M2XKdVYxKM7:TR?= MlnxNVIhcA>? NWLlOlFW[XWpbXXueJMhfGinIHPhd5Bie2VvMzDhZ5Rqfmm2eR?= NH\aSGU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkizO|Y5QSd-MkK4N|c3QDl:L3G+
K562 NFrE[YtHfW6ldHnvckBCe3OjeR?= Ml:wOlAh|ryP NHzXPWsyOiCq MXLheYdu\W62czD0bIUh[2G|cHHz[U0{KGGldHn2bZR6 Mom4QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjJ6M{e2PFkoRjJ{OEO3Olg6RC:jPh?=
HL60/Adr MWjGeY5kfGmxbjDBd5NigQ>? M2PKbFYxKM7:TR?= NVfBbFFnOTJiaB?= M4TZc4F2\22nboTzJJRp\SClYYPwZZNmNTNiYXP0bZZqfHl? NIW2XHY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkizO|Y5QSd-MkK4N|c3QDl:L3G+
HL60 M2L3UGZ2dmO2aX;uJGF{e2G7 NYL6TWU6PjBizszN NFHSO3oyOiCq MUHheYdu\W62czD0bIUh[2G|cHHz[U0{KGGldHn2bZR6 MnfYQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjJ6M{e2PFkoRjJ{OEO3Olg6RC:jPh?=
K562/Adr  MnrYRZBweHSxc3nzJGF{e2G7 NWX4UXUzPjBizszN M{O4TlEzKGh? NWX3O3F1\W6qYX7j[ZMhe2ira3;ubY4ucW6mdXPl[EBieG:ydH;zbZM> NEWyV3o9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkizO|Y5QSd-MkK4N|c3QDl:L3G+
K562 NWjoOJNzSXCxcITvd4l{KEG|c3H5 MV62NEDPxE1? M3TDV|EzKGh? M2nI[4VvcGGwY3XzJJNpcWuxbnnuMYlv\HWlZXSgZZBweHSxc3nz NXjiT3pDRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK4N|c3QDlpPkKyPFM4Pjh7PD;hQi=>
HL60/Adr NU\SToFsSXCxcITvd4l{KEG|c3H5 NXHpTGl{PjBizszN M4XaSFEzKGh? MoXH[Y5p[W6lZYOgd4hqc2:waX6tbY5lfWOnZDDhdI9xfG:|aYO= NETlcYM9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkizO|Y5QSd-MkK4N|c3QDl:L3G+
HL60 NGPkW4tCeG:ydH;zbZMhSXO|YYm= MnrhOlAh|ryP NHXRcVcyOiCq NHHLUFhmdmijbnPld{B{cGmtb37pck1qdmS3Y3XkJIFxd3C2b4Ppdy=> NGrCZZY9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkizO|Y5QSd-MkK4N|c3QDl:L3G+
SH-EP MX\BdI9xfG:|aYOgRZN{[Xl? M4TE[|Ex6oDLzszNxsA> Ml\pO|LjiImqwrC= NGnQcFVqdmirYnn0d{BKSVBiaX7obYJqfG:{LTDhcoQhVGW6YYT1cZVu[WJvaX7keYNm\CCjcH;weI9{cXN? MYO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjh7MEOyNkc,OjJ6OUCzNlI9N2F-
NIH3T3  NV21SFI3TnWwY4Tpc44hSXO|YYm= MofjNVAwPTBizszN M4rPSFEwOyCq NXj5SZR7[W2nbHnvdoF1\XNiVF7G{tEu\HKrdnXuJINwdXCuZYig[o9zdWG2aX;u MW[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzJ4MU[3O{c,OjN{NkG2O|c9N2F-
HT-22  MnHwSpVv[3Srb36gRZN{[Xl? M{HjO|I2yqEQvF2= NGDtSZMx6oDVM{FCpI1qdg>? NYjINWpRTE2VTx?= MVnpcohq[mm2czDFVmshSWO2aY\heIlwdiCrbnT1Z4VlKGK7IHfseZRidWG2ZR?= NEDOcIE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{OwO|c2Oid-MkOzNFc4PTJ:L3G+
HT-22  MoC1R4VtdCCYaXHibYxqfHliQYPzZZk> MmD0NVDDqM7:TR?= NFPocGMyOsLiaB?= M3Pt[WROW09? MmrQdJJwfGWldIOgZYdicW6|dDDncJV1[W2jdHWtbY5lfWOnZDDj[YxtKGSnYYTo MX28ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzNyN{e1Nkc,OjN|MEe3OVI9N2F-
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Jurkat NHmzenlEgXSxcILveIVkfGm4ZTDhd5NigQ>? MVuzNEB2VQ>? MYKxJIhz NIrDc4VEgXSxcILveIVkfGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IFPIXE1qdmS3Y3XkJI5m[3KxcITvd4l{KGmwIHj1cYFvKEq3cnvheEBk\WyuczDhd5Nme3OnZDDhd{BqdmO{ZXHz[UBqdiClZXzsJJZq[WKrbHn0fUBifCB|MDD1UUBqdmO3YnH0[YQh\m:{IEGgbJIh\m:ubH;3[YQh[nliQ1jYJJN1cW23bHH0bY9vKGK7IIDoZZNmKGOxboTyZZN1KG2rY4Lvd4NweHl? NX35bnE{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkm1OFE{PTdpPkK5OVQyOzV5PD;hQi=>
Jurkat MWXDfZRweHKxdHXjeIl3\SCjc4PhfS=> MVizNEB2VQ>? M1;FNVEhcHJ? M4XHRmN6fG:ycn;0[YN1cX[nIHHjeIl3cXS7IHHnZYlve3RiWj3WRWQucW6mdXPl[EBv\WO{b4D0c5NqeyCrbjDoeY1idiCMdYLrZZQh[2WubIOgZZN{\XO|ZXSgZZMhcW6lcnXhd4UhcW5iY3XscEB3cWGkaXzpeJkh[XRiM{CgeW0hcW6ldXLheIVlKG[xcjCxJIhzKG[xbHzve4VlKGK7IGqtWmFFKHO2aX31cIF1cW:wIHL5JJBp[XOnIHPvcpRz[XO2IH3pZ5Jwe2OxcIm= MXy8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQTV2MUO1O{c,Ojl3NEGzOVc9N2F-
Assay
Methods Test Index PMID
Immunofluorescence RIP1 / RIP3 30462730

Protocol (from reference)

Kinase Assay:

[1]

  • RIP1 kinase assay:

    Phosphorylation of RIP1 requires its kinase activity. Expression constructs of FLAGtagged wild-type (WT) or a kinase-inactive pointmutant of RIP1 (K45M) are are transfected into 293T cells and RIP1 kinase assay is performed as described in the Methods in the presence of [γ-32P]ATP for 30 min at 30℃. Samples are subjected to SDS-PAGE and RIP1 band is visualized by autoradiography. Relative intensities of radioactive bands are quantified and are shown (ratio) in this and all other autoradiographs. In parallel to kinase reactions, a sample of beads is subjected to western blot analysis using anti-RIP1 antibody to ensure equal protein amounts in kinase reactions.

Cell Research:

[2]

  • Cell lines: Jurkat, BALB/c 3T3, SV40-transformed MEF, L929
  • Concentrations: 0.01-100 μM
  • Incubation Time: --
  • Method:

    Cells are seeded in 96-well plates (white plates for luminescent assays; black plates for fluorescent assays; clear plates for MTT assay) at the density of 5,000-10,000 cells per well for adherent cells or 20,000-50,000 cells per well for suspension cells in 100 μl of the appropriate phenol red-free media. After incubation, we determined cell viability using one of the following methods. For the ATP assay, we used luminescence-based commercial kits and analyzed luminescence using a Wallac Victor II plate reader. For Sytox assay, we incubated cells with 1 μM Sytox Green reagent for 30 min at 37℃, and then performed fluorescent reading. Subsequently, we added 5 μl of 20% Triton X-100 solution into each well to produce maximal lysis and incubated cells for 1 h at 37℃, then performed the second reading. We calculated the ratio of values before and after Triton treatment and normalized it to the relevant controls not subjected to cytotoxic stimuli, as indicated in figure legends. For the MTT assay, we used the CellTiter 96 AQueous Non-Radioactive Cell Proliferation Assay kit. For PI exclusion assays, we added 2 μg/ml PI into the medium and immediately analyzed samples using FACSCalibur. For PI-annexin V assay we used the ApoAlert Annexin V-EGFP Apoptosis Kit. For DioC6 staining, we incubated cells with 40 nM DiOC6 for 30 min at 37 ℃, washed once and analyzed in FACSCalibur. For ROS analysis, we incubated cells with 5 μM dihydroethidium for 30 min at 37 ℃, washed once and analyzed in FACSCalibur. EM analyses are performed at the Harvard Medical School EM facility. We acquired bright-field images of the cells using an Axiovert 200 microscope.

  • (Only for Reference)

Solubility (25°C)

In vitro

DMSO 51 mg/mL warmed
(196.66 mM)
Water Insoluble
Ethanol Insoluble

In vivo

Add solvents to the product individually and in order
(Data is from Selleck tests instead of citations):
5% DMSO+45% PEG 300+ddH2O
For best results, use promptly after mixing.

10mg/mL

* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.

Chemical Information

Molecular Weight 259.33
Formula

C13H13N3OS

CAS No. 4311-88-0
Storage 3 years -20°C powder
2 years -80°C in solvent
Smiles CN1C(=O)C(NC1=S)CC2=CNC3=CC=CC=C32

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such
as vortex, ultrasound or hot water bath can be used to aid dissolving.

Molarity Calculator

Mass Concentration Volume Molecular Weight

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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