Home Apoptosis RIP kinase inhibitor Necrostatin-1 (Nec-1)

연구용

Necrostatin-1 (Nec-1) RIPK1 Inhibitor

제품 번호: S8037

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 (Nec-1) RIP kinase inhibitor Chemical Structure

화학 구조

분자량: 259.33

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품질 관리 (Quality Control)

배치: 순도: 99.97%
99.97

세포 배양, 처리 및 작업 농도
(Cell Culture, Treatment & Working Concentration)

세포주 분석 유형 농도 배양 시간 제형 활성 설명 PMID
HT-22 Cytotoxicity Assay 10 μM 12 h protects against cell death induced by 5 mmol/L glutamate  17760869
Jurkat  Function Assay 200 μm 30 min reduces Naegleria fowleri-induced reactive oxygen species (ROS) generation 21535020
Jurkat  Cytotoxicity Assay 50/ 100/200 μm 1/3 h reduces Naegleria fowleri-induced cytotoxicity 21535020
SW13 Cell Viability Assay 100 μM 24 h DMSO increases cellular survival 22136818
8505c Cell Viability Assay 100 μM 24 h DMSO increases cellular survival 22136818
TPC-1 Cell Viability Assay 100 μM 24 h DMSO increases cellular survival 22136818
L929sA Apoptosis Assay 10 μM 1 h abrogates the interaction of caspase-8 with FADD 22362767
L929sA Apoptosis Assay 10 μM 1 h rescues cells expressing RIPK1ΔID from TNF-induced apoptosis 22362767
L929sA Apoptosis Assay 10 μM 1 h inhibits the apoptotic response to TNF 22362767
K562/Adr  Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
K562 Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
HL60/Adr Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
HL60 Function Assay 60 μM 12 h increases the activity of caspases, caspase 8 and 9 22837689
K562/Adr  Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
K562 Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
HL60/Adr Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
HL60 Function Assay 60 μM 12 h augments the caspase-3 activity 22837689
K562/Adr  Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
K562 Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
HL60/Adr Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
HL60 Apoptosis Assay 60 μM 12 h enhances shikonin-induced apoptosis 22837689
SH-EP Apoptosis Assay 10 μM  72 h  inhibits IAP inhibitor- and Lexatumumab-induced apoptosis 22890322
NIH3T3  Function Assay 10/50 μM 1/3 h ameliorates TNFα-driven complex formation 23261677
HT-22  Function Assay 25 μM 0–30 min DMSO inhibits ERK Activation induced by glutamate 23307752
HT-22  Cell Viability Assay 10 μM 12 h DMSO protects against glutamate-induced cell death 23307752
KMS-12-BM Cytotoxicity Assay 90 µM 1 h blocks BAY 11-7082 induced rapid cell swelling 23527154
MM.1S  Cytotoxicity Assay 90 µM 1 h blocks BAY 11-7082 induced rapid cell swelling 23527154
ΔN-Karpas 299  Cytotoxicity Assay 20 μM 16 h inhibits CD30-induced cell death 23545938
MEFs Function Assay 20 μM 1/2/4 h suppresses TNFα-induced RIPK1 phosphorylation 23727581
MEFs Cytotoxicity Assay 2/6/20 μM 18 h inhibits TNFα-induced cell death in RelA KO MEFs 23727581
RMS13 Cell Viability Assay 40 μg/ml  24 h rescues GX15-070-induced loss of cell viability 23744296
TE671 Cell Viability Assay 40 μg/ml  24 h rescues GX15-070-induced loss of cell viability 23744296
U87 Function Assay 1 mmol/L 1.5-3 h suppresses the expression of RIP-1 caused by shikonin 23840441
C6 Function Assay 1 mmol/L 1.5-3 h suppresses the expression of RIP-1 caused by shikonin 23840441
U87 Cytotoxicity Assay 1 mmol/L 3 h blocks shikonin induced necrosis 23840441
C6 Cytotoxicity Assay 1 mmol/L 3 h blocks shikonin induced necrosis 23840441
U87 Cell Viability Assay 1 mmol/L 3 h attenuates Shikonin induced glioma cell death 23840441
C6 Cell Viability Assay 1 mmol/L 3 h attenuates Shikonin induced glioma cell death 23840441
L929 Function Assay 5 μg/ml 24 h blocks zVAD induced necroptosis and autophagy 23941769
L929 Function Assay 2 μg/ml  24 h promots caspase-6 (p20) activity and procaspase-6 cleavage 23941769
L929 Growth Inhibition Assay 2/5 μg/ml  24 h reverses the cell growth inhibition and cell death induced by TNFα alone as well as TNFα + zVAD 23941769
L929 Function Assay 2/5 μg/ml  24 h reversed the autophagy induced by TNFα alone as well as TNFα + zVAD 23941769
NRK-52E  Cell Viability Assay 20 μM 24 h inhibits increased Drp1 protein expression after TNF-α Stimulation and ATP Depletion 24351845
NRK-52E  Cell Viability Assay 20 μM 24 h increases cell viability after TNF-α Stimulation and ATP Depletion 24351845
NRK-52E  Cell Viability Assay 20 μM 24 h protects cells from cell death caused by ischemia injury 24351845
AGS Cell Viability Assay 60 μm 1 h prevents shikonin-induced cell death 24463199
L-540  Cell Viability Assay 60 μm 1 h reduces the Givinostat/Sorafenib-induced cell death 24561519
L-540  Function Assay 60 μm 1 h prevents the mitochondrial membrane depolarization 24561519
L-540  Function Assay 60 μm 1 h prevents the generation of ROS 24561519
SK-Hep1 Function Assay 60 μM  18 h blocks β-lapachone-mediated PAR accumulation and AIF translocation to the cytosol  24832602
SK-Hep1 Function Assay 60 μM  18 h inhibits β-Lapachone-induced leakage of HMGB-1  24832602
SK-Hep1 Function Assay 60 μM  18 h blocks β-lapachone-induced morphological change, cell death and PI uptake 24832602
OHC Function Assay 300 μM DMSO increases the number of apoptotic OHCs without altering the levels of CC8 after noise exposure 24874734
OHC Function Assay 300 μM DMSO diminishes noise-induced AMPK activation 24874734
OHC Function Assay 300 μM DMSO results in a reduction of noise-induced RIP1 and RIP3 immunofluorescence 24874734
OHC Function Assay 300 μM DMSO decreases noise-induced swollen nuclei  24874734
OHC Function Assay 300 μM DMSO increases noise-induced condensed nuclei 24874734
Huh7 Cell Viability Assay 50 µM 24/48 h DMSO prevents cell death of rAdHCV co-infected Huh7 cells 24973240
L929 Cell Viability Assay 30 μM 1 h inhibits TNF-α-induced cleavage of Topo I 25095742
L929 Cell Viability Assay 30 μM 1 h inhibits TNF-α-induced loss of cell viability 25095742
L929-A Function Assay 50 μM  12 h inhibits the TNFα-induced loss of mitochondrial membrane permeability 25398540
L929 Function Assay 50 μM  12 h inhibits TNFα-induced Bid cleavage 25398540
L929-N  Function Assay 50 μM  12 h blocks the cleavage of Caspase-3 and PARP 25398540
L929-A  Function Assay 50 μM  12 h blocks the cleavage of Caspase-3 and PARP 25398540
L929-N  Cell Viability Assay 50 μM  24 h blocks TNFα-induced cell death 25398540
L929-A  Cell Viability Assay 50 μM  24 h blocks TNFα-induced cell death 25398540
KMS-12-PE  Cell Viability Assay 60 μM 5 h inhibits SHK-induced cell death 25530098
SGC-7901 Cell Viability Assay 30 μM 1 h suppresses oxaliplatin-mediated cell death 25767076
BxPC-3 Function Assay 20 μM 24 h decreases the early necrotic cells 26000607
MiaPaCa-2 Function Assay 20 μM 24 h decreases the early necrotic cells 26000607
NCI-H28 Cell Viability Assay 10 μM 24 h prevents DAPE-induced reduction of NCI-H28 cell viability  26004138
BMDM  Function Assay 10 μM 30 min protects cells from TAKI-induced LDH release 26381601
MEFs  Cell Viability Assay 10 μM 48 h DMSO inhibits zVAD-promoted death of CNOT3-depleted MEFs 26437789
A549 Cell Viability Assay 50 μM 24 h inhibits MMS-induced cell death 26472723
Jurkat T Necroptosis assay Inhibition of TNF-alpha-induced necroptosis in FADD-deficient human Jurkat T cells, EC50 = 0.05 μM. 18467094
Jurkat Function assay Inhibition of endogenous RIP1 autophosphorylation in human Jurkat cells, EC50 = 0.182 μM. 18408713
Sf9 Function assay 30 mins Inhibition of recombinant human GST-fused RIPK1 (1 to 497 residues) expressed in baculovirus infected insect Sf9 cells in presence of 32P-gamma-ATP after 30 mins by autoradiogram-based Western blot method, IC50 = 0.182 μM. 28280261
Jurkat T Necroptosis assay Effective concentration required for inhibition of necroptosis in FADD deficient Jurkat T cells treated with TNF-alpha, EC50 = 0.49 μM. 16153840
Jurkat Necroptosis assay Inhibition of cellular necroptosis in TNFalpha treated FADD deficient human Jurkat cells, EC50 = 0.49 μM. 18408713
Jurkat T Necroptosis assay 30 uM 24 hrs Inhibition of necroptosis in TNF-alpha-induced human Jurkat T cells assessed as cell viability at 30 uM after 24 hrs 18467094
L929 Necroptosis assay 30 uM 24 hrs Inhibition of necroptosis in zVAD-induced mouse L929 cells assessed as cell viability at 30 uM after 24 hrs 18467094
L929 Necroptosis assay 30 uM 24 hrs Inhibition of necroptosis in TNF-alpha-induced mouse L929 cells assessed as cell viability at 30 uM after 24 hrs 18467094
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necrotic cell death in mouse 3T3 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of TNFalpha and zVAD.fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necrotic cell death in mouse 3T3 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of FasL and zVAD.fmk 16408008
MEF Cell death assay 16 hrs Inhibition of death receptor signaling mediated necrotic cell death in SV40 transformed mouse MEF cells assessed as cell viability after 16 hrs by ATP based viability assay in presence of TNFalpha, CHX and zVAD.fmk 16408008
IEC18 Cell death assay Inhibition of death receptor signaling mediated necrotic cell death in rat IEC18 cells assessed as cell viability in presence of TNFalpha and zVAD.fmk 16408008
HL60 Cell death assay Inhibition of death receptor signaling mediated necrotic cell death in human HL60 cells assessed as cell viability in presence of TNFalpha and zVAD.fmk 16408008
L929 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necrotic cell death in mouse L929 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as nuclear condensation by bright field microscopy in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as organelle swelling by bright field microscopy in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as early loss of plasma membrane integrity by bright field microscopy in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells assessed as appearance of translucent cytosol in presence of FasL, CHX and zVAD-fmk 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of nuclear condensation by bright field microscopy in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of organelle swelling by bright field microscopy in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of early loss of plasma membrane integrity by bright field microscopy in presence of TNFalpha 16408008
Jurkat Necrosis assay Inhibition of necrosis in human Jurkat cells deficient in FADD assessed as inhibition of appearance of translucent cytosol in presence of TNFalpha 16408008
U937 Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human U937 cells assessed as cell viability after 48 hrs by ATP based viability assay in presence of TNFalpha and zVAD-fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as cell viability after 24 hrs by ATP based viability assay in presence of TNFalpha and zVAD-fmk 16408008
Jurkat Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD assessed as decreased levels of PE-conjugated LC3-II (autophagy marker) after 24 hrs by Western blot method in presence of TNFalpha 16408008
L929 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse L929 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of TNFalpha 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of TNFalpha and zVAD-fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of FasL and zVAD-fmk 16408008
3T3 Cell death assay 24 hrs Inhibition of death receptor signaling mediated necroptotic cell death in mouse 3T3 cells assessed as decreased levels of PE-conjugated autophagy marker LC3-II after 24 hrs by Western blot method in presence of rapamycin 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing FKBP12-based dimerization domain assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP K45M mutant assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase domain assessed as cell viability after 48 hrs by FACS in presence of AP1510, zVAD-fmk 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing FKBP12-based dimerization domain assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP K45M mutant assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Jurkat Cell death assay 48 hrs Inhibition of death receptor signaling mediated necroptotic cell death in human Jurkat cells deficient in FADD and expressing RIP kinase domain assessed as cell viability after 48 hrs by FACS in presence of AP1510 16408008
Sf9 Function assay Inhibition of human RIP1 K45M mutant autophosphorylation expressed in Sf9 cells 18408713
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against FasL-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by FasL stimulation measured after 20 hrs by Alamar blue assay 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against CHX-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by CHX stimulation by Alamar blue assay 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against Z-VAD-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by Z-VAD stimulation by Alamar blue assay 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against FasL-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by FasL stimulation measured after 20 hrs by phase contrast microscopy 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against CHX-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by CHX stimulation by phase contrast microscopy 29541357
Jurkat Cytoprotective assay 30 uM 1 hr Cytoprotective activity against Z-VAD-induced necroptosis in human Jurkat cells assessed as increase in cell viability at 30 uM incubated for 1 hr followed by Z-VAD stimulation by phase contrast microscopy 29541357
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화학 정보, 보관 및 안정성 (Chemical Information, Storage & Stability)

분자량 259.33 화학식

C13H13N3OS

 보관 (수령일로부터)
CAS 번호 4311-88-0 SDF 다운로드 원액 보관

동의어 Nec-1 Smiles CN1C(=O)C(NC1=S)CC2=CNC3=CC=CC=C32

용해도 (Solubility)

In vitro
배치:

DMSO : 57 mg/mL (219.79 mM)
(수분으로 오염된 DMSO는 용해도를 감소시킬 수 있습니다. 신선하고 무수 DMSO를 사용하십시오.)

Water : Insoluble

Ethanol : Insoluble

몰농도 계산기

질량 농도 부피 분자량
희석 계산기 분자량 계산기

In vivo
배치:

생체 내 제형 계산기 (투명한 용액)

1단계: 아래 정보 입력 (권장: 실험 중 손실을 고려하여 추가 동물 포함)

mg/kg g μL

2단계: 생체 내 제형 입력 (이것은 계산기일 뿐 제형이 아닙니다. 용해도 섹션에 생체 내 제형이 없는 경우 먼저 당사에 문의하십시오.)

% DMSO % % Tween 80 % ddH2O
%DMSO %

계산 결과:

작업 농도: mg/ml;

DMSO 원액 준비 방법: mg 약물 사전 용해 μL DMSO ( 원액 농도 mg/mL, 농도가 해당 약물 배치의 DMSO 용해도를 초과하는 경우 먼저 당사에 문의하십시오. )

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가μL PEG300, 혼합하고 투명하게 한 다음 추가μL Tween 80, 혼합하고 투명하게 한 다음 추가 μL ddH2O, 혼합하고 투명하게 합니다.

생체 내 제형 준비 방법: 취하다 μL DMSO 원액, 다음 추가 μL 옥수수 기름, 혼합하고 투명하게 합니다.

참고: 1. 다음 용매를 추가하기 전에 액체가 투명한지 확인하십시오.
2. 용매를 순서대로 추가해야 합니다. 다음 용매를 추가하기 전에 이전 추가에서 얻은 용액이 투명한 용액인지 확인해야 합니다. 와동, 초음파 또는 뜨거운 물 중탕과 같은 물리적 방법을 사용하여 용해를 도울 수 있습니다.

작용 메커니즘 (Mechanism of Action)

특징
A powerful tool for characterizing the role of necroptosis with characterized primary target.
Targets/IC50/Ki
IDO
RIP1
(293T cells)
490 nM(EC50)
시험관 내(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 this compound.

This chemical efficiently suppresses necroptotic cell death triggered by an array of stimuli in a variety of cell types. It, 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.

키나아제 분석
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.
생체 내(In vivo)

Necrostatin-1 (Nec-1) is a specific small molecule inhibitor of receptor-interacting protein kinase 1 (RIPK1) that specifically inhibits phosphorylation of this compound.
참조

적용 분야 (Applications)

방법 바이오마커 이미지 PMID
Immunofluorescence RIP1 / RIP3
S8037-IF1
30462730