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Necrostatin-1 (Nec-1) RIPK1-remmer

Name: Necrostatin-1 (Nec-1)
Brand: Selleck Chemicals
SKU: S8037
Rating: 5 (317 reviews)

Cat.Nr.S8037

Necrostatin-1 (Nec-1) is een specifieke RIP1 (RIPK1)-remmer en remt TNF-α-geïnduceerde necroptose met een EC50 van 490 nM in 293T-cellen. Necrostatin-1 blokkeert ook IDO en onderdrukt autophagy en apoptosis.

Necrostatin-1 (Nec-1) RIP kinase Remmer Chemical Structure

Chemische structuur

Moleculair gewicht: 259.33

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Kwaliteitscontrole (Quality Control)

Batch: Zuiverheid: 99.97%

99.97

Gerelateerde doelwitten	Bcl-2 Caspase PD-1/PD-L1 Ferroptosis p53 Apoptosis related Synthetic Lethality STAT TNF-alpha Ras
Overige RIP kinase Inhibitoren	Necrostatin 2 racemate (Nec-1s) GSK872 Mito-TEMPO GSK'963 RIPA-56 GSK583 GSK2982772 Resibufogenin HS-1371 GSK2983559 (compound 3)

Celkweek, behandeling & werkconcentratie
(Cell Culture, Treatment & Working Concentration)

Cellijnen	Assaytype	Concentratie	Incubatietijd	Formulering	Activiteitsbeschrijving	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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Chemische informatie, Opslag en Stabiliteit (Chemical Information, Storage & Stability)

Moleculair gewicht	259.33	Formule	C₁₃H₁₃N₃OS	Opslag (Vanaf de ontvangstdatum)	3 jaar-20°Cpoeder
CAS-nr.	4311-88-0	SDF downloaden		Opslag van stamoplossingen	1 jaar-80°Cin oplosmiddel 1 maand-20°Cin oplosmiddel
Synoniemen	Nec-1			Smiles	CN1C(=O)C(NC1=S)CC2=CNC3=CC=CC=C32

Oplosbaarheid (Solubility)

In vitro
Batch:

DMSO : 57 mg/mL (219.79 mM)
(Met vocht verontreinigde DMSO kan de oplosbaarheid verminderen. Gebruik verse, watervrije DMSO.)

Water : Insoluble

Ethanol : Insoluble

Molariteitscalculator

Massa	Concentratie	Volume	Moleculair gewicht
=	×	×

Verdunningscalculator Moleculair gewicht calculator

In vivo Batch:
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Clear solution	5%DMSO 1 Corn oil Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	2.850mg/ml (10.99mM)	Taking the 1 mL working solution as an example, add 50 μL of 57 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Clear solution	5%DMSO 1 10%PEG 300 2 85 %ddH2O Gevalideerd door Selleck labs. Mocht u aanpassingen aan deze formulering nodig hebben, neem dan contact op met ons verkoopteam voor aangepaste tests.	1.200mg/ml (4.63mM)	Taking the 1 mL working solution as an example, add 50 μL of 24 mg/ml clarified DMSO stock solution to 100 μL of PEG300, mix evenly to clarify it; add 850 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.
Homogeneous suspension	CMC-NA	≥5mg/ml	Taking the 1 mL working solution as an example, add 5 mg of this product to 1 ml of CMC-Na solution, mix evenly to obtain a homogeneous suspension with a final concentration of 5 mg/ml.

In vivo Formuleringscalculator (Heldere oplossing)

Stap 1: Voer de onderstaande informatie in (Aanbevolen: Een extra dier voor het geval van verlies tijdens het experiment)

Dosering: mg/kg Gemiddeld gewicht van dieren: g Doseervolume per dier:μL Aantal dieren:

Stap 2: Voer de in vivo formulering in (Dit is alleen de calculator, geen formulering. Neem eerst contact met ons op als er geen in vivo formulering is in het gedeelte Oplosbaarheid.)

% DMSO + % + % Tween 80 + % ddH₂O

%DMSO+ %

Berekeningsresultaten:

Werkconcentratie: mg/ml;

Methode voor het bereiden van DMSO-mastervloeistof: mg geneesmiddel vooraf opgelost in μL DMSO ( Concentratie mastervloeistof mg/mL, Neem eerst contact met ons op als de concentratie de DMSO-oplosbaarheid van de partij geneesmiddel overschrijdt. )

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toeμL PEG300, mengen en helder maken, voeg vervolgens toeμL Tween 80, mengen en helder maken, voeg vervolgens toe μL ddH₂O, mengen en helder maken.

Methode voor het bereiden van in vivo formulering: Neem μL DMSO mastervloeistof, voeg vervolgens toe μL Maïsolie, mengen en helder maken.

Opmerking: 1. Zorg ervoor dat de vloeistof helder is voordat u het volgende oplosmiddel toevoegt.
2. Zorg ervoor dat u het/de oplosmiddel(en) in de juiste volgorde toevoegt. U moet ervoor zorgen dat de verkregen oplossing, bij de vorige toevoeging, een heldere oplossing is voordat u verdergaat met het toevoegen van het volgende oplosmiddel. Fysische methoden zoals vortexen, echografie of een warmwaterbad kunnen worden gebruikt om het oplossen te bevorderen.

Werkingsmechanisme (Mechanism of Action)

Kenmerken	A powerful tool for characterizing the role of necroptosis with characterized primary target.
Targets/IC₅₀/Ki	IDO RIP1 (293T cells) 490 nM(EC50)
In vitro	Necrostatin-1 (1-100 μM) remt de autofosforylering van overgeëxpresseerd en endogeen RIP1. Er is vastgesteld dat RIP1 het primaire cellulaire doelwit is dat verantwoordelijk is voor de antinecroptose-activiteit van deze verbinding. Deze chemische stof onderdrukt efficiënt necroptotische celdood, veroorzaakt door een reeks stimuli in verschillende celtypen. Het, eerder geïdentificeerd als een kleinmoleculige remmer van necroptose, remt RIP kinase-geïnduceerde necroptose en remt TNF-α-geïnduceerde necroptose in Jurkat-cellen met een EC50 van 490 nM.
Kinase Assay	RIP1-kinase-assay
Kinase Assay	Fosforylering van RIP1 vereist de kinase-activiteit ervan. Expressieconstructies van FLAG-getagd wildtype (WT) of een kinase-inactieve puntmutant van RIP1 (K45M) worden getransfecteerd in 293T-cellen en de RIP1 kinase-assay wordt uitgevoerd zoals beschreven in de methoden in aanwezigheid van [γ-³²P]ATP gedurende 30 min bij 30℃. Monsters worden onderworpen aan SDS-PAGE en de RIP1-band wordt gevisualiseerd door autoradiografie. Relatieve intensiteiten van radioactieve banden worden gekwantificeerd en worden (verhouding) getoond in deze en alle andere autoradiografieën. Parallel aan kinase-reacties wordt een monster van kralen onderworpen aan western blot-analyse met anti-RIP1-antilichaam om gelijke eiwithoeveelheden in kinase-reacties te garanderen.
In vivo	Necrostatin-1 (Nec-1) is een specifieke kleine moleculaire remmer van receptor-interacterend proteïnekinase 1 (RIPK1) die specifiek de fosforylering van deze verbinding remt.
Referenties	[1] http://www.ncbi.nlm.nih.gov/pubmed/?term=18408713 [2] http://www.ncbi.nlm.nih.gov/pubmed/?term=16408008 [3] https://pubmed.ncbi.nlm.nih.gov/30542285/

Toepassingen (Applications)

Methoden	Biomarkers	Afbeeldingen	PMID
Immunofluorescence	RIP1 / RIP3		30462730

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C5=C4/X	C5:	LOG(C5):
C6=C5/X	C6:	LOG(C6):
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C8=C7/X	C8:	LOG(C8):