Necrostatin-1

Name: Necrostatin-1
Brand: Selleck Chemicals
SKU: S8037
Rating: 5 (91 reviews)

For research use only.

Catalog No.S8037 Synonyms: Nec-1

91 publications

CAS No. 4311-88-0

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.

Selleck's Necrostatin-1 has been cited by 91 publications

Haematologica, 2021, 10.3324/haematol.2020.259531

PubMed: 33406814 ( click the link to review the publication )

Stem Cell Reports, 2021, S2213-6711(21)00158-2

PubMed: 33891868 ( click the link to review the publication )

Cancers (Basel), 2021, 13(2)E243

PubMed: 33440739 ( click the link to review the publication )

FASEB J, 2021, 35(5):e21561

PubMed: 33864423 ( click the link to review the publication )

J Mol Cell Cardiol, 2021, 150:65-76

PubMed: 33098823 ( click the link to review the publication )

Front Cell Dev Biol, 2021, 9:651360

PubMed: 33842478 ( click the link to review the publication )

Biomolecules, 2021, 11(2)153

PubMed: 33503908 ( click the link to review the publication )

Int J Mol Sci, 2021, 22(5)2702

PubMed: 33800107 ( click the link to review the publication )

Toxicol In Vitro, 2021, 73:105146

PubMed: 33737050 ( click the link to review the publication )

Autophagy, 2021, 1-15

PubMed: 33629929 ( click the link to review the publication )

Immunohorizons, 2021, 5(2):90-101

PubMed: 33597177 ( click the link to review the publication )

Transl Stroke Res, 2021, 10.1007/s12975-021-00888-3

PubMed: 33629276 ( click the link to review the publication )

Mol Neurobiol, 2021, 10.1007/s12035-021-02320-1

PubMed: 33634378 ( click the link to review the publication )

Sci Adv, 2020, 25;6(13):eaay9789

PubMed: 32232155 ( click the link to review the publication )

Part Fibre Toxicol, 2020, 17(1):46

PubMed: 32948194 ( click the link to review the publication )

Redox Biol, 2020, 38:101777

PubMed: 33166869 ( click the link to review the publication )

J Med Chem, 2020, 63(17):9838-9855

PubMed: 32809827 ( click the link to review the publication )

Cancers (Basel), 2020, 12(11)E3205

PubMed: 33143299 ( click the link to review the publication )

Cell Death Dis, 2020, 11(9):781

PubMed: 32951003 ( click the link to review the publication )

Cell Death Dis, 2020, 22;11(5):391

PubMed: 32444644 ( click the link to review the publication )

Mol Nutr Food Res, 2020, 1;e1901231

PubMed: 32476238 ( click the link to review the publication )

Oxid Med Cell Longev, 2020, 2020:1295382

PubMed: 33123307 ( click the link to review the publication )

Oxid Med Cell Longev, 2020, 4;2020:5021983

PubMed: 32322335 ( click the link to review the publication )

Oncogenesis, 2020, 9(10):94

PubMed: 33093450 ( click the link to review the publication )
Int J Mol Sci, 2020, 21(12)E4204

PubMed: 32545631 ( click the link to review the publication )

J Biol Chem, 2020, 1;295(18):6236-6248

PubMed: 32179652 ( click the link to review the publication )

Acta Pharmacol Sin, 2020, 10.1038/s41401-020-0376-8

PubMed: 32132657 ( click the link to review the publication )

Front Pharmacol, 2020, 11:812

PubMed: 32595496 ( click the link to review the publication )

Biochem Biophys Res Commun, 2020, 8;521(2):414-419

PubMed: 31672277 ( click the link to review the publication )

Med Sci Monit, 2020, 26:e919739

PubMed: 32019905 ( click the link to review the publication )

Biosci Biotechnol Biochem, 2020, 18;1-8

PubMed: 32419644 ( click the link to review the publication )

Sci Total Environ, 2020, 699:134296

PubMed: 31683218 ( click the link to review the publication )

Cell Death Dis, 2020, 11(8):635

PubMed: 32801338 ( click the link to review the publication )

Cell Rep, 2020, 30(1):25-36

PubMed: 31914391 ( click the link to review the publication )

Cell Death Discov, 2020, 6:67

PubMed: 32793397 ( click the link to review the publication )

STAR Protoc, 2020, 1(1)100034

PubMed: 32793892 ( click the link to review the publication )

PLoS Pathog, 2020, 16(7):e1008473

PubMed: 32649716 ( click the link to review the publication )

Blood Adv, 2020, 4(22):5666-5680

PubMed: 33211827 ( click the link to review the publication )

Cell Death Dis, 2020, 11(8):630

PubMed: 32801360 ( click the link to review the publication )

Int J Oncol, 2020, 56(2):439-447

PubMed: 31894265 ( click the link to review the publication )

J Clin Invest, 2019, 129(6):2431-2445

PubMed: 31081803 ( click the link to review the publication )

EMBO Rep, 2019, 20(10):e47840

PubMed: 31410978 ( click the link to review the publication )

Cell Death Differ, 2019, 10.1038/s41418-019-0274-0

PubMed: 30760873 ( click the link to review the publication )

Front Oncol, 2019, 9:436

PubMed: 31192135 ( click the link to review the publication )

Sci Rep, 2019, 9(1):2902

PubMed: 30814594 ( click the link to review the publication )

J Biol Chem, 2019, 10.1074/jbc.RA119.010379

PubMed: 31723031 ( click the link to review the publication )

Front Mol Neurosci, 2019, 12:222

PubMed: 31607859 ( click the link to review the publication )

Int J Mol Med, 2019, 43(2):967-979

PubMed: 30569134 ( click the link to review the publication )
Cell Transplant, 2019, 28(11):1358-1372

PubMed: 31370690 ( click the link to review the publication )

Oncotarget, 2019, 10(62):6678-6690

PubMed: 31803362 ( click the link to review the publication )

Mol Metab, 2019, 23:14-23

PubMed: 30837196 ( click the link to review the publication )

J Hepatol, 2018, 68(6):1228-1238

PubMed: 29408314 ( click the link to review the publication )

Nat Commun, 2018, 9(1):1680

PubMed: 29700283 ( click the link to review the publication )

Autophagy, 2018, 14(5):778-795

PubMed: 29171784 ( click the link to review the publication )

Clin Cancer Res, 2018, 24(23):6066-6077

PubMed: 30061362 ( click the link to review the publication )

Elife, 2018, 10.7554/eLife.40167

PubMed: 30561326 ( click the link to review the publication )

Oncogene, 2018, 37(29):4033-4045

PubMed: 29706658 ( click the link to review the publication )

Oncoimmunology, 2018, 7(6):e1431086

PubMed: 29872558 ( click the link to review the publication )

Mol Oncol, 2018, 10.1002/1878-0261.12422

PubMed: 30537101 ( click the link to review the publication )

Biochim Biophys Acta Mol Cell Res, 2018, 1865(3):522-531

PubMed: 29278689 ( click the link to review the publication )

J Immunol, 2018, 200(8):2826-2834

PubMed: 29563176 ( click the link to review the publication )

Am J Cancer Res, 2018, 8(8):1514-1527

PubMed: 30210920 ( click the link to review the publication )

Ann Biomed Eng, 2018, 46(7):1013-1025

PubMed: 29603044 ( click the link to review the publication )

Toxicology, 2018, 406-407:33-43

PubMed: 29860048 ( click the link to review the publication )

Am J Transl Res, 2018, 10(5):1357-1372

PubMed: 29887951 ( click the link to review the publication )

Int J Mol Med, 2018, 41(5):3051-3062

PubMed: 29436589 ( click the link to review the publication )

Int J Med Sci, 2018, 15(14):1648-1657

PubMed: 30588188 ( click the link to review the publication )

Cancer Commun (Lond), 2018, 38(1):43

PubMed: 29970185 ( click the link to review the publication )

Cancer Lett, 2017, 381(1):96-103

PubMed: 27477897 ( click the link to review the publication )

Cancer Lett, 2017, 402:81-92

PubMed: 28576750 ( click the link to review the publication )

Cell Death Dis, 2017, 8(5):e2765

PubMed: 28471454 ( click the link to review the publication )

Genes Chromosomes Cancer, 2017,

PubMed: 28177558 ( click the link to review the publication )
Cell Cycle, 2017, 16(4):348-359

PubMed: 27929749 ( click the link to review the publication )

Nat Commun, 2016, 7:10274

PubMed: 26817517 ( click the link to review the publication )

Autophagy, 2016, 12(4):659-70

PubMed: 27050456 ( click the link to review the publication )

Autophagy, 2016, 12(10):1738-1758

PubMed: 27463126 ( click the link to review the publication )

Cancer Res, 2016, 10.1158/0008-5472.CAN-16-0523

PubMed: 27302164 ( click the link to review the publication )

J Exp Clin Cancer Res, 2016, 8;35(1):192

PubMed: 27931237 ( click the link to review the publication )

Free Radical Biology and Medicine, 2016, S0891-5849(16)00114-3

PubMed: 27012422 ( click the link to review the publication )

Sci Rep, 2016, 6:24245

PubMed: 27095146 ( click the link to review the publication )

Oncol Rep, 2016, 36(2):1135-42

PubMed: 27349281 ( click the link to review the publication )

Oncotarget, 2016, 7(30):47387-47402

PubMed: 27329722 ( click the link to review the publication )

Oncotarget, 2016,

PubMed: 27014915 ( click the link to review the publication )

Oncotarget, 2016, 7(25):38718-38730

PubMed: 27231850 ( click the link to review the publication )

Mutat Res, 2016, 789:1-8

PubMed: 27155559 ( click the link to review the publication )

Genet Mol Res, 2016, 10.4238/gmr.15027298

PubMed: 27323061 ( click the link to review the publication )

Cancer Res, 2015, 10.1158/0008-5472.CAN-14-2199

PubMed: 25724678 ( click the link to review the publication )

J Cell Mol Med, 2015, 10.1111/jcmm.12495

PubMed: 25754842 ( click the link to review the publication )

PLoS One, 2015, 10(8):e0134144

PubMed: 26270345 ( click the link to review the publication )

PLoS One, 2015, 10(3):e0122083

PubMed: 25894744 ( click the link to review the publication )

Oncotarget, 2015, 6(15):13476-86

PubMed: 25974963 ( click the link to review the publication )

Purity & Quality Control

Choose Selective RIP kinase Inhibitors

Biological Activity

Description

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	NFzCeJhEgXSxdH;4bYNqfHliQYPzZZk>	MkfnNVAh\|ryP	NX7uW4ZmOTJiaB?=		MWXwdo91\WO2czDh[4FqdnO2IHPlcIwh\GWjdHigbY5lfWOnZDDifUA2yqCvbX;sM2wh\2y3dHHtZZRmyqB?	M{\uflxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF5N{[wPFY6Lz5zN{e2NFg3QTxxYU6=
Jurkat	M1z3XWZ2dmO2aX;uJGF{e2G7	NH7RRmMzODEEoN88cS=>	NFK0SmQ{OCCvaX6=		NWnkSpVsemWmdXPld:KhVmGnZ3zldoliKG[xd3zldokucW6mdXPl[EBz\WGldHn2[UBwgHmpZX6gd5Bm[2mnczCoVm9UMSCpZX7ldoF1cW:w	MofiQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjF3M{WwNlAoRjJzNUO1NFIxRC:jPh?=
Jurkat	NIfNXohEgXSxdH;4bYNqfHliQYPzZZk>	MlXaOVAwKDFyMD:yNFDDqM7:bR?=	NWroeIs5OS9\|IHi=		MX3y[YR2[2W\|wrDOZYVodGW{aXGg[o94dGW{aT3pcoR2[2WmIHP5eI91d3irY3n0fS=>	MkPUQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjF3M{WwNlAoRjJzNUO1NFIxRC:jPh?=
SW13	M1v2bWNmdGxiVnnhZoltcXS7IFHzd4F6	MWWxNFAh\|ryP	NEnndo4zPCCq	NHnEU5JFVVOR	MWrpcoNz\WG\|ZYOgZ4VtdHWuYYKgd5Vzfmm4YXy=	NFfqVIw9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkGzOlgyQCd-MkKxN\|Y5OTh:L3G+
8505c	MWjD[YxtKF[rYXLpcIl1gSCDc4PhfS=>	MWKxNFAh\|ryP	MmfTNlQhcA>?	M3;md2ROW09?	NHzIfG5qdmO{ZXHz[ZMh[2WubIXsZZIhe3W{dnn2ZYw>	MlHTQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjJzM{[4NVgoRjJ{MUO2PFE5RC:jPh?=
TPC-1	MlXvR4VtdCCYaXHibYxqfHliQYPzZZk>	MoPmNVAxKM7:TR?=	M3fqbVI1KGh?	MVTEUXNQ	Ml\abY5kemWjc3XzJINmdGy3bHHyJJN2en[rdnHs	NELGUW49[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkGzOlgyQCd-MkKxN\|Y5OTh:L3G+
L929sA	M{TvSWFxd3C2b4Ppd{BCe3OjeR?=	M3HSWFExKM7:TR?=	NFTU[WgyKGh?		M3nrWIFjem:pYYTld{B1cGViaX70[ZJi[3Srb36gc4Yh[2G\|cHHz[U05KHerdHigSmFFTA>?	NXTxSG9rRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkKzOlI4PjdpPkKyN\|YzPzZ5PD;hQi=>
L929sA	MnWzRZBweHSxc3nzJGF{e2G7	MkD4NVAh\|ryP	Mne5NUBp		MknxdoV{[3WnczDj[YxteyCneIDy[ZN{cW6pIGLJVGsy\|pSLRDDmdo9uKFSQRj3pcoR2[2WmIHHwc5B1d3Orcx?=	MWW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjN4Mke2O{c,OjJ\|NkK3Olc9N2F-
L929sA	M2XQe2Fxd3C2b4Ppd{BCe3OjeR?=	M2LBdFExKM7:TR?=	NYj3clJzOSCq		NWnWU3o{cW6qaXLpeJMhfGinIHHwc5B1d3SrYzDy[ZNxd26\|ZTD0c{BVVkZ?	NVOxRYh2RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkKzOlI4PjdpPkKyN\|YzPzZ5PD;hQi=>
K562/Adr	NVrBcm5NTnWwY4Tpc44hSXO\|YYm=	NGjUPZg3OCEQvF2=	NVvON2NjOTJiaB?=		NGfNcYVqdmO{ZXHz[ZMhfGinIHHjeIl3cXS7IH;mJINie3Cjc3XzMEBk[XOyYYPlJFgh[W6mIEm=	MYm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjh\|N{[4PUc,OjJ6M{e2PFk9N2F-
K562	M1TFXWZ2dmO2aX;uJGF{e2G7	MV:2NEDPxE1?	NVHr[lZmOTJiaB?=		MVLpcoNz\WG\|ZYOgeIhmKGGldHn2bZR6KG:oIHPhd5Bie2W\|LDDjZZNx[XOnIEigZY5lKDl?	MkjHQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjJ6M{e2PFkoRjJ{OEO3Olg6RC:jPh?=
HL60/Adr	MnLrSpVv[3Srb36gRZN{[Xl?	NXfBNpJwPjBizszN	MWKxNkBp		MWXpcoNz\WG\|ZYOgeIhmKGGldHn2bZR6KG:oIHPhd5Bie2W\|LDDjZZNx[XOnIEigZY5lKDl?	M{TWV\|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{OEO3Olg6Lz5{MkizO\|Y5QTxxYU6=
HL60	Mmj2SpVv[3Srb36gRZN{[Xl?	NIXZ[JA3OCEQvF2=	MYmxNkBp		NFXPNHhqdmO{ZXHz[ZMhfGinIHHjeIl3cXS7IH;mJINie3Cjc3XzMEBk[XOyYYPlJFgh[W6mIEm=	NGjOenI9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkizO\|Y5QSd-MkK4N\|c3QDl:L3G+
K562/Adr	M4C4WGZ2dmO2aX;uJGF{e2G7	M4DuO\|YxKM7:TR?=	NEKwbXMyOiCq		Mk\tZZVodWWwdIOgeIhmKGOjc4Dhd4UuOyCjY4Tpeol1gQ>?	M3f0eVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{OEO3Olg6Lz5{MkizO\|Y5QTxxYU6=
K562	NELTRVVHfW6ldHnvckBCe3OjeR?=	MmXoOlAh\|ryP	M1i4XVEzKGh?		NFnBXXpifWevZX70d{B1cGViY3HzdIF{\S1\|IHHjeIl3cXS7	MkjyQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjJ6M{e2PFkoRjJ{OEO3Olg6RC:jPh?=
HL60/Adr	M4f0dGZ2dmO2aX;uJGF{e2G7	NWC4dWNSPjBizszN	NGD0[ngyOiCq		MXTheYdu\W62czD0bIUh[2G\|cHHz[U0{KGGldHn2bZR6	NYjTWmNFRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK4N\|c3QDlpPkKyPFM4Pjh7PD;hQi=>
HL60	Mnj1SpVv[3Srb36gRZN{[Xl?	M3n2S\|YxKM7:TR?=	NHLocWUyOiCq		Mn7mZZVodWWwdIOgeIhmKGOjc4Dhd4UuOyCjY4Tpeol1gQ>?	M3PUT\|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{OEO3Olg6Lz5{MkizO\|Y5QTxxYU6=
K562/Adr	MWPBdI9xfG:\|aYOgRZN{[Xl?	NGO4ZVc3OCEQvF2=	Mnj3NVIhcA>?		Ml2w[Y5p[W6lZYOgd4hqc2:waX6tbY5lfWOnZDDhdI9xfG:\|aYO=	MXW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjh\|N{[4PUc,OjJ6M{e2PFk9N2F-
K562	NVvvWGJpSXCxcITvd4l{KEG\|c3H5	M1vYR\|YxKM7:TR?=	NV\UW4EzOTJiaB?=		NHX4d5VmdmijbnPld{B{cGmtb37pck1qdmS3Y3XkJIFxd3C2b4Ppdy=>	MWG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjh\|N{[4PUc,OjJ6M{e2PFk9N2F-
HL60/Adr	M2CwemFxd3C2b4Ppd{BCe3OjeR?=	NULJcWlwPjBizszN	MkLuNVIhcA>?		NYnxN5Zn\W6qYX7j[ZMhe2ira3;ubY4ucW6mdXPl[EBieG:ydH;zbZM>	NVvE[2UzRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK4N\|c3QDlpPkKyPFM4Pjh7PD;hQi=>
HL60	NFjwWWNCeG:ydH;zbZMhSXO\|YYm=	NG\4Xm03OCEQvF2=	NVHHWHJrOTJiaB?=		MVTlcohidmOnczDzbIlsd26rbj3pcoR2[2WmIHHwc5B1d3Orcx?=	M3LISlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ{OEO3Olg6Lz5{MkizO\|Y5QTxxYU6=
SH-EP	NVPTR4ZySXCxcITvd4l{KEG\|c3H5	MWKxNQKBkc7:TdMg	M{P0Olcz6oDLaNMg		NWjadVhycW6qaXLpeJMhUUGSIHnubIljcXSxcj2gZY5lKEyneHH0eY12dWGkLXnu[JVk\WRiYYDvdJRwe2m\|	MXG8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOjh7MEOyNkc,OjJ6OUCzNlI9N2F-
NIH3T3	NIjVOWpHfW6ldHnvckBCe3OjeR?=	M3LZblExNzVyIN88US=>	MUSxM\|MhcA>?		Mo\zZY1mdGmxcnH0[ZMhXE6IzsGt[JJqfmWwIHPvcZBt\XhiZn;ycYF1cW:w	NGXJW4E9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{K2NVY4Pyd-MkOyOlE3Pzd:L3G+
HT-22	M1zaTmZ2dmO2aX;uJGF{e2G7	MX2yOeKh\|ryP	M{j1U\|DjiJN\|MNMgcYlv	M1jCSGROW09?	NF3pUJFqdmirYnn0d{BGWktiQXP0bZZifGmxbjDpcoR2[2WmIHL5JIdtfXSjbXH0[S=>	NXvDeIt6RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkOzNFc4PTJpPkKzN\|A4PzV{PD;hQi=>
HT-22	MV7D[YxtKF[rYXLpcIl1gSCDc4PhfS=>	NHfQOGEyOMLizszN	NIOxOI4yOsLiaB?=	MXvEUXNQ	Mnv1dJJwfGWldIOgZYdicW6\|dDDncJV1[W2jdHWtbY5lfWOnZDDj[YxtKGSnYYTo	M1j3ZVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ\|M{C3O\|UzLz5{M{OwO\|c2OjxxYU6=
KMS-12-BM	MmjhR5l1d3SxeHnjbZR6KEG\|c3H5	NHGxW3A6OCEEtV2=	NHzIfY8yKGh?		NGXRdm5jdG:la4OgRmF[KDFzLUewPFIhcW6mdXPl[EBz[XCrZDDj[YxtKHO5ZXzsbY5o	NGDqbpk9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{M{WyO\|E2PCd-MkO1NlcyPTR:L3G+
MM.1S	NVfjT5FvS3m2b4TvfIlkcXS7IFHzd4F6	M{D1fVkxKML3TR?=	MY[xJIg>		NWjwcXF4[myxY3vzJGJCYSBzMT23NFgzKGmwZIXj[YQhemGyaXSgZ4VtdCC\|d3XscIlv\w>?	MnT1QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN3MkexOVQoRjJ\|NUK3NVU1RC:jPh?=
ΔN-Karpas 299	MnvXR5l1d3SxeHnjbZR6KEG\|c3H5	NFXKVlAzOMLizszN	MofXNVYhcA>?		MkfZbY5pcWKrdIOgR2Q{OC2rbnT1Z4VlKGOnbHyg[IVifGh?	MmDuQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN3NEW5N\|goRjJ\|NUS1PVM5RC:jPh?=
MEFs	MmPjSpVv[3Srb36gRZN{[Xl?	NVfobohCOjEEoN88US=>	MlHPNU8zNzRiaB?=		MnTVd5VxeHKnc4Pld{BVVkcQsT3pcoR2[2WmIGLJVGsyKHCqb4PwbI9zgWyjdHnvci=>	M4nCN\|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ\|N{K3OVgyLz5{M{eyO\|U5OTxxYU6=
MEFs	NFLEWJdEgXSxdH;4bYNqfHliQYPzZZk>	Ml\RNk83NzJyIN88US=>	M{PNfVE5KGh?		NUTVU3hlcW6qaXLpeJMhXE6IzsGtbY5lfWOnZDDj[YxtKGSnYYToJIlvKFKnbFGgT28hVUWIcx?=	NUi3bWlHRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkO3Nlc2QDFpPkKzO\|I4PThzPD;hQi=>
RMS13	MmixR4VtdCCYaXHibYxqfHliQYPzZZk>	MUK0NQKBkc7:Zz;tcOKh	MVuyOEBp		M2LnPJJme2O3ZYOgS3gyPS1yN{CtbY5lfWOnZDDsc5N{KG:oIHPlcIwhfmmjYnnsbZR6	MXy8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzd2NEK5Okc,OjN5NESyPVY9N2F-
TE671	Mo\uR4VtdCCYaXHibYxqfHliQYPzZZk>	MWm0NQKBkc7:Zz;tcOKh	MnSxNlQhcA>?		MWry[ZNkfWW\|IFfYNVUuODdyLXnu[JVk\WRibH;zd{Bw\iClZXzsJJZq[WKrbHn0fS=>	M4LGPFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ\|N{S0Nlk3Lz5{M{e0OFI6PjxxYU6=
U87	M1TVdWZ2dmO2aX;uJGF{e2G7	NHnae3gyKG2vb3yvUC=>	NE\4eIQyNjVvMzDo		MkHHd5VxeHKnc4Pld{B1cGViZYjwdoV{e2mxbjDv[kBTUVBvMTDjZZV{\WRiYomgd4hqc2:waX6=	MmXSQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN6NEC0OFEoRjJ\|OESwOFQyRC:jPh?=
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U87	NHjUTWxEgXSxdH;4bYNqfHliQYPzZZk>	NV3xUos4OSCvbX;sM2w>	NETyNmQ{KGh?		MoTnZoxw[2u\|IIPobYtwdmmwIHnu[JVk\WRibnXjdo9{cXN?	MWe8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzh2MES0NUc,OjN6NEC0OFE9N2F-
C6	MYrDfZRwfG:6aXPpeJkhSXO\|YYm=	Moe2NUBudW:uL1y=	MXGzJIg>		MVPicI9kc3Nic3jpb49vcW5iaX7keYNm\CCwZXPyc5Nqew>?	NWTKRllZRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkO4OFA1PDFpPkKzPFQxPDRzPD;hQi=>
U87	MY\D[YxtKF[rYXLpcIl1gSCDc4PhfS=>	MoTuNUBudW:uL1y=	NGnEW5c{KGh?		MUHheJRmdnWjdHXzJHNpcWuxbnnuJIlv\HWlZXSg[4xqd22jIHPlcIwh\GWjdHi=	MWO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzh2MES0NUc,OjN6NEC0OFE9N2F-
C6	M{nXbGNmdGxiVnnhZoltcXS7IFHzd4F6	NVT4[Wd2OSCvbX;sM2w>	NYTyOXRSOyCq		NYf5UJFi[XS2ZX71ZZRmeyCVaHnrc45qdiCrbnT1Z4VlKGeuaX;tZUBk\WyuIHTlZZRp	MXm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzh2MES0NUc,OjN6NEC0OFE9N2F-
L929	NIXveWNHfW6ldHnvckBCe3OjeR?=	MoDJOeKh\|rypL33s	Mn7FNlTDqGh?		NFXR[oFjdG:la4OgfnZCTCCrbnT1Z4VlKG6nY4LvdJRwe2m\|IHHu[EBifXSxcHjh[5k>	Ml\UQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjN7NEG3OlkoRjJ\|OUSxO\|Y6RC:jPh?=
L929	NYLDeop2TnWwY4Tpc44hSXO\|YYm=	M1vJSlLDqM7:Zz;tcOKh	M1zuPVI1yqCq		MnSzdJJwdW:2czDjZZNx[XOnLU[gLJAzOCliYXP0bZZqfHliYX7kJJBzd2Ojc4Dhd4UuPiClbHXheoFo\Q>?	MUm8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzl2MUe2PUc,OjN7NEG3Olk9N2F-
L929	M4jlZmdzd3e2aDDJcohq[mm2aX;uJGF{e2G7	MY[yM\|XDqM7:Zz;tcOKh	M1LOV\|I1yqCq		NWTtS4syemW4ZYLz[ZMhfGinIHPlcIwh\3Kxd4ToJIlvcGmkaYTpc44h[W6mIHPlcIwh\GWjdHigbY5lfWOnZDDifUBVVkcQsTDhcI9v\SCjczD3[YxtKGG\|IGTOSu6yyqBtwrD6WmFF	MX68ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzl2MUe2PUc,OjN7NEG3Olk9N2F-
L929	NYLZO5JrTnWwY4Tpc44hSXO\|YYm=	NWjHbIFoOi93wrFOwIcwdW{EoB?=	M1HhSVI1yqCq		M1mzWZJmfmW{c3XkJJRp\SCjdYTvdIhi\3liaX7keYNm\CCkeTDUUmbPuSCjbH;u[UBieyC5ZXzsJIF{KFSQRt8xxsAsyqC8VlHE	MWq8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOzl2MUe2PUc,OjN7NEG3Olk9N2F-
NRK-52E	MV3D[YxtKF[rYXLpcIl1gSCDc4PhfS=>	M2fvclIxKM7:TR?=	MmezNlQhcA>?		M1\uSolvcGmkaYTzJIlv[3KnYYPl[EBFenBzIIDyc5RmcW5iZYjwdoV{e2mxbjDh[pRmeiCWTl[t{tEhW3SrbYXsZZRqd25iYX7kJGFVWCCGZYDs[ZRqd25?	NX:zSlNRRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkSzOVE5PDVpPkK0N\|UyQDR3PD;hQi=>
NRK-52E	MoPnR4VtdCCYaXHibYxqfHliQYPzZZk>	MXKyNEDPxE1?	NWPiN4o6OjRiaB?=		MnWybY5kemWjc3XzJINmdGxidnnhZoltcXS7IHHmeIVzKFSQRj5OtUBUfGmvdXzheIlwdiCjbnSgRXRRKESncHzleIlwdg>?	NHzhT3Q9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEO1NVg1PSd-MkSzOVE5PDV:L3G+
NRK-52E	NFLNb4tE\WyuIG\pZYJqdGm2eTDBd5NigQ>?	MmSxNlAh\|ryP	MX:yOEBp		M2DsTJBzd3SnY4TzJINmdGy\|IH\yc40h[2WubDDk[YF1cCClYYXz[YQh[nliaYPjbIVucWFiaX7qeZJ6	M{nidFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2M{WxPFQ2Lz5{NEO1NVg1PTxxYU6=
AGS	NYXrXohPS2WubDDWbYFjcWyrdImgRZN{[Xl?	Ml;TOlDjiIoQvH2=	MlK3NgKBkWh?		NHfWTJRxemW4ZX70d{B{cGmtb37pck1qdmS3Y3XkJINmdGxiZHXheIg>	M1K2VFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2NE[zNVk6Lz5{NES2N\|E6QTxxYU6=
L-540	MVXD[YxtKF[rYXLpcIl1gSCDc4PhfS=>	NH\1coo3OOLCid88cS=>	MkjxNgKBkWh?		NVzv[WFLemWmdXPld{B1cGViR3n2bY5we3SjdD;Tc5Ji\mWwaXKtbY5lfWOnZDDj[YxtKGSnYYTo	MYO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDV4MUWxPUc,OjR3NkG1NVk9N2F-
L-540	M{LqPGZ2dmO2aX;uJGF{e2G7	M2LrclYx6oDLzszt	NW[4SHNWOeLCiXi=		NFjRZYxxemW4ZX70d{B1cGVibXn0c4Npd26mcnnhcEBu\W2kcnHu[UBl\XCxbHHybZpifGmxbh?=	NXL4OmRSRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS1OlE2OTlpPkK0OVYyPTF7PD;hQi=>
L-540	M2DWTmZ2dmO2aX;uJGF{e2G7	NHzzdnM3OOLCid88cS=>	MWmx5qCKcA>?		MVvwdoV3\W62czD0bIUh\2WwZYLheIlwdiCxZjDSU3M>	NIPsbVM9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NEW2NVUyQSd-MkS1OlE2OTl:L3G+
SK-Hep1	MXzGeY5kfGmxbjDBd5NigQ>?	MnjQOlDjiIoQvF5CpC=>	NVTHT3ZqOTkkgJno		Mn\KZoxw[2u\|wrFOtk1t[XCjY3jvcoUudWWmaXH0[YQhWEGUIHHjZ5VufWyjdHnvckBidmRiQVnGJJRz[W6\|bH;jZZRqd25idH:geIhmKGO7dH;zc4zDqA>?	NYPsfINXRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS4N\|I3ODJpPkK0PFMzPjB{PD;hQi=>
SK-Hep1	NGTjbZZHfW6ldHnvckBCe3OjeR?=	NV7r[YFEPjEkgJpOwG3DqA>?	NXqycYY6OTkkgJno		MlG2bY5pcWKrdIOg{tIuVGGyYXPoc45mNWmwZIXj[YQhdGWja3Hn[UBw\iCKTVfCMVHDqA>?	NXTmR\|F{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS4N\|I3ODJpPkK0PFMzPjB{PD;hQi=>
SK-Hep1	MmTzSpVv[3Srb36gRZN{[Xl?	NV;H[5NUPjEkgJpOwG3DqA>?	MX6xPQKBkWh?		M1va[oJtd2Otc9Mg{tIudGGyYXPoc45mNWmwZIXj[YQhdW:{cHjvcI9ocWOjbDDjbIFv\2VuIHPlcIwh\GWjdHigZY5lKFCLIIXweIFs\Q>?	MoP6QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR6M{K2NFIoRjJ2OEOyOlAzRC:jPh?=
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OHC	NFjDWYZHfW6ldHnvckBCe3OjeR?=	MmfSN\|Ax6oDLzszN		M2\kRmROW09?	M1vKeZJme3WudIOgbY4h[SC{ZXT1Z5Rqd25ib3[gco9qe2VvaX7keYNm\CCUSWCxJIFv\CCUSWCzJIludXWwb3\seY9z\XOlZX7j[S=>	NXXTd4VxRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS4O\|Q4OzRpPkK0PFc1PzN2PD;hQi=>
OHC	M3zSd2Z2dmO2aX;uJGF{e2G7	MYSzNFDjiIoQvF2=		NX3aPFJLTE2VTx?=	NETTUGpl\WO{ZXHz[ZMhdm:rc3WtbY5lfWOnZDDze49tdGWwIH71Z4xmccLi	M{\mflxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ2OEe0O\|M1Lz5{NEi3OFc{PDxxYU6=
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Huh7	NI\ZfItE\WyuIG\pZYJqdGm2eTDBd5NigQ>?	MWS1NEDDvU1?	NX;me5A1OjRxNEigbC=>	NHzHcIlFVVOR	MkS2dJJmfmWwdIOgZ4VtdCCmZXH0bEBw\iC{QXTIR3Yh[29vaX7m[YN1\WRiSIXoO{Bk\Wyucx?=	MWW8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDl5M{K0NEc,OjR7N{OyOFA9N2F-
L929	NHeyU4RE\WyuIG\pZYJqdGm2eTDBd5NigQ>?	NYTHTYpMOzEEoN88US=>	NFXacJkyyqCq		NHrxN5ZqdmirYnn0d{BVVkZvzsGtbY5lfWOnZDDjcIVifmGpZTDv[kBVd3CxIFm=	NX;RNpg{RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkWwPVU4PDJpPkK1NFk2PzR{PD;hQi=>
L929	MYnD[YxtKF[rYXLpcIl1gSCDc4PhfS=>	NIP5RlE{OMLizszN	M{HiblHDqGh?		M{TWfolvcGmkaYTzJHRPTi4QsT3pcoR2[2WmIHzvd5Mhd2ZiY3XscEB3cWGkaXzpeJk>	NVHOcFRVRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkWwPVU4PDJpPkK1NFk2PzR{PD;hQi=>
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Jurkat	NFjjVVFP\WO{b4Ppd{Bie3OjeR?=				M1LCTmlvcGmkaYTpc44hd2ZibnXjdo9{cXNiaX6gbJVu[W5iSoXyb4F1KGOnbHzzJIRm\mmlaXXueEBqdiCIQVTEJIF{e2W\|c3XkJIF{KGmwaHnibZRqd25ib3[g[YFzdHlibH;zd{Bw\iCybHHzcYEhdWWvYoLhcoUhcW62ZXfybZR6KGK7IHLybYdpfCCoaXXs[EBucWO{b4Pjc5B6KGmwIIDy[ZNmdmOnIH;mJHRPTmGucHjh	M3TXdVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF4NEC4NFA5Lz5zNkSwPFAxQDxxYU6=
Jurkat	M4W3[W5m[3Kxc3nzJIF{e2G7				M13SOWlvcGmkaYTpc44hd2ZibnXjdo9{cXNiaX6gbJVu[W5iSoXyb4F1KGOnbHzzJIRm\mmlaXXueEBqdiCIQVTEJIF{e2W\|c3XkJIF{KGmwaHnibZRqd25ib3[gZZBx\WG{YX7j[UBw\iC2cnHud4x2[2WwdDDjfZRwe2:uIHnuJJBz\XOnbnPlJI9nKFSQRnHsdIhi	M4D1O\|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzF4NEC4NFA5Lz5zNkSwPFAxQDxxYU6=
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Jurkat	M3nxR2NmdGxiZHXheIgh[XO\|YYm=		MXWyOEBpenN?		MVvJcohq[mm2aX;uJI9nKGSnYYToJJJm[2WydH;yJJNq\26jbHnu[{Bu\WSrYYTl[EBv\WO{b4D0c5Rq[yClZXzsJIRm[XSqIHnuJIh2dWGwIFr1dotifCClZXzsd{Bl\W[rY3nlcpQhcW5iRlHESEBie3Onc4Pl[EBieyCmZXPy[YF{\WRibHX2[Yx{KG:oIGDFMYNwdmq3Z3H0[YQhVEN\|LVnJJEhifXSxcHjh[5khdWG{a3XyLUBi\nSncjCyOEBpenNiYomgW4V{fGW{bjDicI91KG2ndHjv[EBqdiCycnXz[Y5k\SCxZjDUUmZidHCqYR?=	NYfYToNJRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMU[0NFgxODhpPkG2OFA5ODB6PD;hQi=>
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Jurkat	NHzDV2NE\WyuIHTlZZRpKGG\|c3H5		NXrC[phSPDhiaILz		M4XORWlvcGmkaYTpc44hd2ZiZHXheIghemWlZYD0c5Ihe2mpbnHsbY5oKG2nZHnheIVlKG6nY4LvdJRwfGmlIHPlcIwh\GWjdHigbY4hcHWvYX6gTpVzc2G2IHPlcIx{KGSnZnnjbYVvfCCrbjDGRWRFKGGwZDDlfJBz\XO\|aX7nJHJKWCCtaX7hd4Uh[XO\|ZYPz[YQh[XNiY3XscEB3cWGkaXzpeJkh[W[2ZYKgOFghcHK\|IHL5JGZCS1NiaX6gdJJme2WwY3Wgc4YhSVBzNUGwMEB7XkGGLX\tby=>	MmL6QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOTZ2MEiwNFgoRjF4NEC4NFA5RC:jPh?=
Jurkat	M2jscmNmdGxiZHXheIgh[XO\|YYm=		M3XsTFQ5KGi{cx?=		M4n0eGlvcGmkaYTpc44hd2ZiZHXheIghemWlZYD0c5Ihe2mpbnHsbY5oKG2nZHnheIVlKG6nY4LvdJRwfGmlIHPlcIwh\GWjdHigbY4hcHWvYX6gTpVzc2G2IHPlcIx{KGSnZnnjbYVvfCCrbjDGRWRFKGGwZDDlfJBz\XO\|aX7nJHJKWCCNNEXNJI12fGGwdDDhd5Nme3OnZDDhd{Bk\WyuII\pZYJqdGm2eTDh[pRmeiB2ODDodpMh[nliRlHDV{BqdiCycnXz[Y5k\SCxZjDBVFE2OTBuIIrWRWQu\m2t	NHPuXYc9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9zNkSwPFAxQCd-MU[0NFgxODh:L3G+
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Jurkat	M4TiZ2NmdGxiZHXheIgh[XO\|YYm=		NHS3WIY1QCCqcoO=		MVHJcohq[mm2aX;uJI9nKGSnYYToJJJm[2WydH;yJJNq\26jbHnu[{Bu\WSrYYTl[EBv\WO{b4D0c5Rq[yClZXzsJIRm[XSqIHnuJIh2dWGwIFr1dotifCClZXzsd{Bl\W[rY3nlcpQhcW5iRlHESEBidmRiZYjwdoV{e2mwZzDSTXAhc2mwYYPlJIF{e2W\|c3XkJIF{KGOnbHygeoli[mmuaYT5JIFnfGW{IES4JIhzeyCkeTDGRWNUKGmwIIDy[ZNmdmOnIH;mJGFROTVzMB?=	NVrIUIJ1RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC\|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMU[0NFgxODhpPkG2OFA5ODB6PD;hQi=>
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Jurkat	NFLqXpREgXSxcILveIVkfGm4ZTDhd5NigQ>?	NHHLe3A{OCC3TR?=	MWWxJIhz		NHXXXIxEgXSxcILveIVkfGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IGqtWmFFNWmwZIXj[YQhdmWlcn;weI9{cXNiaX6gbJVu[W5iSoXyb4F1KGOnbHzzJIF{e2W\|c3XkJIF{KGmwY4LlZZNmKGmwIHPlcIwhfmmjYnnsbZR6KGG2IEOwJJVOKGmwY4XiZZRm\CCob4KgNUBpeiCob3zsc5dm\CCkeTDaMXZCTCC\|dHnteYxifGmxbjDifUBCdGGvYYKgZox2\SCjc4PhfS=>	NHK2OmU9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OUW0NVM2Pyd-Mkm1OFE{PTd:L3G+
Jurkat	M{DRPGN6fG:ycn;0[YN1cX[nIHHzd4F6	MUizNEB2VQ>?	NI\DWGQyKGi{		NVe3WG5[S3m2b4Dyc5Rm[3SrdnWgZYN1cX[rdImgZYdicW6\|dDDGZZNNNWmwZIXj[YQhdmWlcn;weI9{cXNiaX6gbJVu[W5iSoXyb4F1KGOnbHzzJIF{e2W\|c3XkJIF{KGmwY4LlZZNmKGmwIHPlcIwhfmmjYnnsbZR6KGG2IEOwJJVOKGmwY4XiZZRm\CCob4KgNUBpeiCob3zsc5dm\CCkeTDGZZNNKHO2aX31cIF1cW:wIH3lZZN2emWmIHHmeIVzKDJyIHjyd{BjgSCyaHHz[UBkd262cnHzeEBucWO{b4Pjc5B6	NIfXWm89[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OUW0NVM2Pyd-Mkm1OFE{PTd:L3G+
Jurkat	NH7DU49EgXSxcILveIVkfGm4ZTDhd5NigQ>?	MmDaN\|AhfU1?	MmTSNUBpeg>?		NGnNcXJEgXSxcILveIVkfGm4ZTDhZ5Rqfmm2eTDh[4FqdnO2IFPIXE1qdmS3Y3XkJI5m[3KxcITvd4l{KGmwIHj1cYFvKEq3cnvheEBk\WyuczDhd5Nme3OnZDDhd{BqdmO{ZXHz[UBqdiClZXzsJJZq[WKrbHn0fUBifCB\|MDD1UUBqdmO3YnH0[YQh\m:{IEGgbJIh\m:ubH;3[YQh[nliQ1jYJJN1cW23bHH0bY9vKGK7IIDoZZNmKGOxboTyZZN1KG2rY4Lvd4NweHl?	M3HJUVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7NUSxN\|U4Lz5{OUW0NVM2PzxxYU6=
Jurkat	NFfLSolEgXSxcILveIVkfGm4ZTDhd5NigQ>?	MXuzNEB2VQ>?	MUSxJIhz		MlrQR5l1d3C{b4TlZ5RqfmViYXP0bZZqfHliYXfhbY5{fCCcLW\BSE1qdmS3Y3XkJI5m[3KxcITvd4l{KGmwIHj1cYFvKEq3cnvheEBk\WyuczDhd5Nme3OnZDDhd{BqdmO{ZXHz[UBqdiClZXzsJJZq[WKrbHn0fUBifCB\|MDD1UUBqdmO3YnH0[YQh\m:{IEGgbJIh\m:ubH;3[YQh[nliWj3WRWQhe3SrbYXsZZRqd25iYomgdIhie2ViY3;ueJJie3RibXnjdo9{[2:yeR?=	M{DkR\|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7NUSxN\|U4Lz5{OUW0NVM2PzxxYU6=

... Click to View More Cell Line Experimental Data

Assay

Methods	Test Index	PMID
Immunofluorescence	RIP1 / RIP3 ; PubMed: 30462730 PLoS One Immunofluorscence data showed an increased expression of RIP1 and RIP3 in Pg-LPS group and an eminent reduction of RIP1 in the Pg-LPS+Nec-1 group.	30462730

Protocol

Kinase Assay:

^[1]

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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]

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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)

References

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 Download Necrostatin-1 SDF

Molecular Weight	259.33
Formula	C₁₃H₁₃N₃OS
CAS No.	4311-88-0
Storage	3 years-20°C powder 2 years-80°C in solvent
Synonyms	Nec-1
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)

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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

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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.

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.

Bio Calculators

Molarity Calculator

Calculate the mass, volume or concentration required for a solution. The Selleck molarity calculator is based on the following equation:

Mass (mg) = Concentration (mM) × Volume (mL) × Molecular Weight (g/mol)

*When preparing stock solutions, please always use the batch-specific molecular weight of the product found on the via label and MSDS / COA (available on product pages).

Dilution Calculator

Calculate the dilution required to prepare a stock solution. The Selleck dilution calculator is based on the following equation:

Concentration _(start) x Volume _(start) = Concentration _(final) x Volume _(final)

This equation is commonly abbreviated as: C1V1 = C2V2 ( Input Output )

* When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / COA (available online).

The Serial Dilution Calculator Equation

Serial Dilutions
Concertration (start):
Dilution-folds:

Computed Result

C1=C0/X	C1:	LOG(C1):
C2=C1/X	C2:	LOG(C2):
C3=C2/X	C3:	LOG(C3):
C4=C3/X	C4:	LOG(C4):
C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
C7=C6/X	C7:	LOG(C7):
C8=C7/X	C8:	LOG(C8):

Molecular Weight Calculator

Enter the chemical formula of a compound to calculate its molar mass and elemental composition:

Tip: Chemical formula is case sensitive. C10H16N2O2 c10h16n2o2

Instructions to calculate molar mass (molecular weight) of a chemical compound:

To calculate molar mass of a chemical compound, please enter its chemical formula and click 'Calculate'.

Definitions of molecular mass, molecular weight, molar mass and molar weight:

Molecular mass (molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.

Molarity Calculator

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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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Necrostatin-1