基本信息

技术规格

物性(参考值)

基本信息

产品描述

Necrostatin-1是一种特异性RIP1 (RIPK1)抑制剂，抑制TNF-α诱导的细胞坏死，在293T细胞中EC50为490 nM。

靶点（IC50 & Targe）

RIP1,490nM(EC50)

体外研究

Necrostatin-1 (1-100 μM) 抑制过表达和内源性的RIP1发生自磷酸化。RIP1是初级细胞靶点，负责Necrostatin-1的抗细胞坏死活性。[1] Necrostatin-1有效抑制多种类型细胞触发的坏死性细胞死亡。Necrostatin-1作为细胞坏死的小分子抑制剂, 作用于jurkat细胞，抑制RIP激酶的诱导细胞坏死，抑制TNF-α诱导的细胞坏死，EC50为490 nM。[2]

激酶实验

RIP1 激酶检测:

RIP1 的磷酸化需要其激酶活性。FLAG标记的野生型(WT)或RIP1(K45M) 突变体失活激酶的表达结构转染到293T细胞中，在有γ-32PATP存在时，RIP1激酶实验在30°C下进行30分钟。样品进行SDS-PAGE，通过放射自显影可观察到RIP1带。对放射性带的相对强度进行量化，并显示比率。在激酶反应的同时，珠样本使用anti-RIP1抗体进行Western Blot分析，确保与激酶反应中等量的蛋白。

细胞实验

Cell lines: Jurkat, BALB/c 3T3, SV40-转化的MEF, L929

Concentrations: 0.01-100 μM

Incubation Time: --

Method:细胞接种在96孔板中（白色板进行发光检测;黑色板进行荧光检测;空白板进行MTT实验）贴壁细胞按每孔5000-10000个细胞的密度接种，悬浮细胞按每孔20,000-50,000个细胞的密度接种，孔中含100 μl合适的无酚红培养基。温育后，使用如下方法之一测定细胞存活率。ATP 实验中，使用购买的发光试剂盒，并使用Wallac Victor II酶标仪分析发光值。Sytox实验中,细胞与 1 μM Sytox Green试剂在37°C下温育30分钟,然后进行荧光读数。随后，增加5 μl 20% Triton X-100溶液到每孔中，产生最大溶解，细胞37°C下温育1小时，然后进行二次读数。Triton处理前和后，计算值的比率。MTT实验，使用CellTiter 96 AQueous 非放射性细胞增殖检测试剂盒。PI排除实验中, 加入2 μg/ml PI 到培养基中，立即使用FACSCalibur分析样品。PI-膜联蛋白V 实验中，使用ApoAlert Annexin V-EGFP 凋亡试剂盒。进行DioC6染色, 细胞与40 nM DiOC6 在37°C下温育30分钟, 洗涤一次，使用FACSCalibur分析。ROS分析中, 细胞与5 μM Dihydroethidium在37°C下温育30分钟, 洗涤一次，使用FACSCalibur分析。

(Only for Reference)

参考文献

[1] Degterev A, et al. Nat Chem Biol, 2008, 4(5), 313-321.

[2] Degterev A, et al. Nat Chem Biol, 2005, 1(2), 112-119.

安全信息

图形或危害标志
提示语	Warning
危险说明	H315 H319 H335
防范说明	P261 P271 P280
WGK	3

产品说明

一般描述

A cell-permeable, potent, and selective blocker of necroptosis (EC₅₀ = 494 nM in FADD-deficient Jurkat cells treated with TNF-α), a nonapoptotic necrotic cell death pathway mediated by death-domain receptors (DRs) that offers neuroprotection in a murine model of ischemic brain injury. Exhibits no effect on DR-induced apoptosis. Also acts as a selective and ATP-competitive inhibitor of RIP1 kinase with negligible effect of RIP2 kinase activity. Nec-1 target appears to be a critical common necroptotic step upstream of execution events and downstream of DRs. Inactive control, Nec-1i, is also available (Cat. No. 480066). Also available as a 25 mM solution in DMSO (Cat. No. 505224).
A cell-permeable, potent, and selective blocker of necroptosis (EC₅₀ = 494 nM in FADD-deficient Jurkat cells treated with TNF-α), a nonapoptotic necrotic cell death pathway mediated by death-domain receptors (DRs) that offers neuroprotection in a murine model of ischemic brain injury. Exhibits no effect on DR-induced apoptosis. Also acts as a selective and ATP-competitive inhibitor of RIP1 kinase with negligible effect of RIP2 kinase activity. Nec-1 target appears to be a critical common necroptotic step upstream of execution events and downstream of DRs. Inactive control, Nec-1i, is also available (Cat. No. 480066).

包装

5 mg in Plastic ampoule
Packaged under inert gas

生化/生理作用

Cell permeable: yes
Primary Target
Blocker of necroptosis
Reversible: no
Product competes with ATP.
EC50 = 494 nM in blocking necrosis in FADD-deficient Jurkat cells treated with TNF-α

警告

Toxicity: Standard Handling (A)

重悬

Following reconstitution, aliquot and freeze (-20°C). Stock solutions are stable for up to 3 months at -20°C.

其他说明

Degterev, A., et al. 2013. Nat. Chem. Biol.9, 192.
Degterev, A., et al. 2012. Cell Death Differ.20, 366.
Christofferson, D.E., et al. 2012. Cell Death Dis.3, e320.
Takahashi, N., et al. 2012. Cell Death Dis.3, e437.
Degterev, A., et al. 2008. Nat. Chem. Biol.4, 313.
Degterev, A., et al. 2005. Nat. Chem. Biol.1, 112.
Muller, A.J., et al. 2005. Nat. Med.11, 312.
Teng, X., et al. 2005. Bioorg. Med. Chem. Lett.15, 5039.

法律信息

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

基本信息

产品性质

安全信息

产品说明

应用

Necrostatin-1已用于研究在小鼠体内肝损伤过程中及在体外肝星状细胞中，其对白介素(IL)-33表达的调节作用。在实验研究中，它还被用作受体相互作用蛋白激酶1(RIPK1)的抑制剂。

生化/生理作用

Necrostatin-1(Nec-1)可能对脑外伤 (TBI)患者具有治疗作用。另外，它还可用于治疗其他细胞死亡方式为神经坏死的急性中枢神经系统(CNS)疾病。它广泛用于研究实验疾病模型中受体相互作用蛋白激酶1 (RIPK1)对细胞死亡和炎症的影响。Nec-1通过提高细胞谷胱甘肽 (GSH)水平、减少活性氧产生，抑制凋亡诱导因子和B细胞淋巴瘤2(Bcl-2)的核转运以及B细胞淋巴瘤2(Bcl-2)/腺病毒E1B 19kDa互作蛋白3相关途径，从而保护海马(HT-22)细胞免受谷氨酸诱导的氧化作用。
Necrostatin-1是坏死性凋亡的抑制剂(非凋亡性细胞死亡途径)。

特点和优势

该化合物是细胞凋亡研究的推荐产品。点击此处，了解更多精选的细胞凋亡产品。了解更多有关用于其他研究领域的生物活性小分子的信息，请访问 sigma.com/discover-bsm。

基本信息

产品性质

安全信息

基本信息

安全信息

图形或危害标志
危险性说明	H315: Causes skin irritation. H319: Causes serious eye irritation. H335: May cause respiratory irritation.
防范说明	P233: Keep container tightly closed. P260: Do not breathe . P261: Avoid breathing . P264: Wash hands thoroughly after handling. P271: Use only outdoors or in a well-ventilated area. P280: Wear . P302 + P352: IF ON SKIN: Wash with plenty of soap and water. P304: IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing.. P304 + P340: IF INHALED: Remove victim to fresh air and keep at rest in a position comfortable for breathing. P305 + P351 + P338: IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing. P312: Call a POISON CENTER or doctor/physician if you feel unwell. P321: Specific treatment (see relevant procedures. on this label). P332 + P313: If skin irritation occurs: Get medical advice/ attention. P337 + P313: If eye irritation persists: Get medical advice/attention. P340: Remove victim to fresh air and keep at rest in a position comfortable for breathing. P362: Take off contaminated clothing and wash before reuse. P403: Store in a well-ventilated place. P403 + P233: Store in a well-ventilated place. Keep container tightly closed. P405: Store locked up. P501: Dispose of contents/container to in accordance with local regulation .