
Biological Activity
Potent, selective, cell-permeable and irreversible inhibitor of phosphatidylinositol 3-kinase (PI 3-kinase) (IC50 = 2 - 4 nM). Also potently inhibits polo-like kinase 1 (PLK1) (IC50 = 5.8 nM).Technical Data
The technical data provided above is for guidance only.For batch specific data refer to the Certificate of Analysis.Tocris products are intended for laboratory research use only, unless stated otherwise.
Background References
- Wortmannin is a potent phophatidylinositol 3-kinase inhibitor: the role of phosphatidylinositol 3,4,5-trisphosphate in neutrophil responses.Arcaro and WymannBiochem.J., 1993;296:297
- Wortmannin, a potent and selective inhibitor of phosphatidylinositol-3-kinase.Powis et al.Cancer Res., 1994;54:2419
- Wortmannin, a widely used phosphoinositide 3-kinase inhibitor, also potently inhibits mammalian polo-like kinase.Liu et al.Chem.Biol., 2005;12:99
- In vitro and in vivo antitumor activity of the phosphatidylinositol-3-kinase inhibitor wortmannin.Schultz et al.Anticancer Res., 1995;15:1135
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Citations for Wortmannin
The citations listed below are publications that use Tocris products.Selected citations for Wortmannin include:
63Citations: Showing 1 - 10
- Pancreatic Ins content regulation by the estrogen receptor ER alpha.Authors: Alonso-Magdalena Et al.PLoS One;3:e2069
- Onion peel extract and its constituent, quercetin inhibits human Slo3 in a pH and calcium dependent manner.Authors: Wijerathne Et al.Korean J Physiol Pharmacol2019;23:381
- The PI3K and MAPK/p38 pathways control stress granule assembly in a hierarchical manner.Authors: Heberle Et al.Life Sci Alliance2019;2
- Age-associated cholesterol reduction triggers brain Ins. resistance by facilitating ligand-independent receptor activation and pathway desensitization.Authors: Martín-Segura Et al.Aging Cell2019;18:e12932
- Integrin CD11b negatively regulates Mincle-induced signaling via the Lyn-SIRPα-SHP1 complex.Authors: Zhang Et al.Exp Mol Med2018;50:e439
- The homozygous CX3CR1-M280 mutation impairs human monocyte survival.Authors: Collar Et al.JCI Insight2018;3
- Initiation of Behavioral Response to Antidepressants by Cholecystokinin Neurons of the Dentate Gyrus.Authors: Medrihan Et al.Neuron2017;95:564
- Induction of Bex genes by curcumin is associated with apoptosis and activation of p53 in N2a neuroblastoma cells.Authors: Sidhar and GiriSci Rep2017;7:41420
- Molecular mechanisms involved in the non-monotonic effect of bisphenol-a on Ca2+ entry in mouse pancreatic β-cells.Authors: Villar-Pazos Et al.Sci Rep2017;7:11770
- Phosphatidylinositol-4,5-bisphosphate is required for KCNQ1/KCNE1 channel function but not anterograde trafficking.Authors: Royal Et al.PLoS One2017;12:e0186293
- PACAP and VIP Inhibit the Invasiveness of Glioblastoma Cells Exposed to Hypoxia through the Regulation of HIFs and EGFR Expression.Authors: Maugeri Et al.Oncotarget2016;7:139
- Mincle-mediated translational regulation is required for strong nitric oxide production and inflammation resolution.Authors: Lee Et al.Oncotarget2016;7:11322
- The GABAB positive allosteric modulators CGP7930 and GS39783 stimulate ERK1/2 signalling in cellslacking functional GABAB receptorsAuthors: Olianas Et al.European Journal of Pharmacology2016;794:135
- SPECC1L deficiency results in increased adherens junction stability and reduced cranial neural crest cell delamination.Authors: Wilson Et al.Sci Rep2016;6:17735
- Effects of novel muscarinic M3 receptor ligand C1213 in pulmonary arterial hypertension models.Authors: AhmedPhysiological Reports2016;4:e13069
- Progesterone receptor membrane component-1 regulates hepcidin biosynthesis.Authors: Li Et al.J Clin Invest2016;126:389
- The LPA1/ZEB1/miR-21-activation pathway regulates metastasis in basal breast cancer.Authors: Sahay Et al.PLoS One2015;6:20604
- Endothelial Rac1 is essential for hematogenous metastasis to the lung.Authors: Yao Et al.Sci Rep2015;6:17501
- Stimulation of EphB2 attenuates tau phosphorylation through PI3K/Akt-mediated inactivation of glycogen synthase kinase-3β.Authors: Jiang Et al.Mol Cancer2015;5:11765
- α6-Containing GABAA Receptors Are the Principal Mediators of Inhibitory Synapse Strengthening by Ins in Cerebellar Granule Cells.Authors: Accardi Et al.J Neurosci2015;35:9676
- Neuregulin 1 signalling modulates mGluR1 function in mesencephalic dopaminergic neurons.Authors: Ledonne Et al.Mol Psychiatry2015;20:959
- Receptor protein tyrosine phosphatase β/ζ is a functional binding partner for vascular endothelial growth factor.Authors: Koutsioumpa Et al.Oncotarget2015;14:19
- Upregulation of Nicotinic Acetylcholine Receptor α4+β2 through a Ligand-Independent PI3Kβ Mechanism That Is Enhanced by TNFα and the Jak2/p38Mapk Pathways.Authors: Rogers and GahringFront Pharmacol2015;10:e0143319
- Regulation of neurons in the dorsal motor nucleus of the vagus by SIRT1.Authors: Jiang and ZsombokFront Neurosci2014;7:270
- Lysophosphatidic acid induces vasodilation mediated by LPA1 receptors, phospholipase C, and endothelial nitric oxide synthase.Authors: Ruisanchez Et al.FASEB J2014;28:880
- Centmitor-1, a novel acridinyl-acetohydrazide, possesses similar molecular interaction field and antimitotic cellular phenotype as rigosertib, on 01910.Na.Authors: Mäki-Jouppila Et al.Nat Commun2014;13:1054
- Mechanosensitive ATP release from hemichannels and Ca2+ influx through TRPC6 accelerate wound closure in keratinocytes.Authors: Takada Et al.J Cell Sci2014;127:4159
- A novel and essential role for FcγRIIa in cancer cell-induced platelet activation.Authors: Mitrugno Et al.Blood2014;123:249
- Regulation of nutrient-sensitive autophagy by uncoordinated 51-like kinases 1 and 2.Authors: McAlpine Et al.Autophagy2013;9:361
- Combination of LC3 shRNA plasmid transfection and genistein treatment inhibited autophagy and increased apoptosis in malignant neuroblastoma in cell culture and animal models.Authors: Mohan Et al.PLoS One2013;8:e78958
- Herpes simplex virus 2 (HSV-2) prevents dendritic cell maturation, induces apoptosis, and triggers release of proinflammatory cytokines: potential links to HSV-HIV synergy.Authors: Stefanidou Et al.Mol Immunol2013;87:1443
- EI24 regulates epithelial-to-mesenchymal transition and tumor progression by suppressing TRAF2-mediated NF-κB activity.Authors: Choi Et al.J Virol2013;4:2383
- An essential role for inhibitor-2 regulation of protein phosphatase-1 in synaptic scaling.Authors: Siddoway Et al.J Pharmacol Exp Ther2013;33:11206
- Melatonin acts through MT1/MT2 receptors to activate hypothalamic Akt and suppress hepatic gluconeogenesis in rats.Authors: Faria Et al.Am J Physiol Endocrinol Metab2013;305:E230
- Cellular Ins resistance disrupts leptin-mediated control of neuronal signaling and transcription.Authors: Nazarians-Armavil Et al.Mol Endocrinol2013;27:990
- HSV activates Akt to trigger calcium release and promote viral entry: novel candidate target for treatment and suppression.Authors: Cheshenko Et al.FASEB J2013;27:2584
- The expression and relaxant effect of bitter taste receptors in human bronchi.Authors: Grassin-Delyle Et al.Respir Res2013;14:134
- The class I PI3K/Akt pathway is critical for cancer cell survival in dogs and offers an opportunity for therapeutic intervention.Authors: Chen Et al.BMC Vet Res2012;8:73
- Long-lasting LTP requires neither repeated trains for its induction nor protein synthesis for its development.Authors: Villers Et al.PLoS One2012;7:e40823
- Endogenous urotensin II selectively modulates erectile function through eNOS.Authors: Bianca Et al.PLoS One2012;7:e31019
- An essential role for modulation of hyperpolarization-activated current in the development of binaural temporal precision.Authors: Khurana Et al.J Neurosci2012;32:2814
- TNF-α triggers rapid membrane insertion of Ca(2+) permeable AMPA receptors into adult motor neurons and enhances their susceptibility to slow excitotoxic injury.Authors: Yin Et al.J Cell Sci2012;238:93
- CDK5 is essential for soluble amyloid β-induced degradation of GKAP and remodeling of the synaptic actin cytoskeleton.Authors: Roselli Et al.PLoS One2011;6:e23097
- Pharmacological and genetic accumulation of hypoxia-inducible factor-1alpha enhances excitatory synaptic transmission in hippocampal neurons through the production of vascular endothelial growth factor.Authors: Huang Et al.J Neurosci2010;30:6080
- FRP inhibits ox-LDL-induced endothelial cell apoptosis through an Akt-NF-{kappa}B-Bcl-2 pathway and inhibits endothelial cell apoptosis in an apoE-knockout mouse model.Authors: Liu Et al.Am J Physiol Endocrinol Metab2010;299:E351
- Disassembly of shank and homer synaptic clusters is driven by soluble beta-amyloid(1-40) through divergent NMDAR-dependent signalling pathways.Authors: Roselli Et al.PLoS One2009;4:e6011
- FcεRI-induced activation by low antigen concentrations results in nuclear signals in the absence of degranulation.Authors: Grodzki Et al.Exp Neurol2009;46:2539
- Inhibitory effect of ETB receptor on Na(+)-K(+) ATPase activity by extracellular Ca(2+) entry and Ca(2+) release from the endoplasmic reticulum in renal proximal tubule cells.Authors: Liu Et al.Hypertens Res2009;32:846
- c-jun controls the ability of IL-12 to induce IL-10 production from human memory CD4+ T cells.Authors: Garcia Et al.J Immunol2009;183:4475
- Kinase networks integrate profiles of N-MthD.-aspartate receptor-mediated gene expression in hippocampus.Authors: Coba Et al.J Biol Chem2008;283:34101
- Antigenic experience dictates functional role of glycogen synthase kinase-3 in human CD4+ T cell responses.Authors: Garcia Et al.J Immunol2008;181:8363
- The novel adipocytokine visfatin exerts direct cardioprotective effects.Authors: Lim Et al.J Cell Mol Med2008;12:1395
- Temporal profiling of changes in phosphatidylinositol 4,5-bisphosphate, inositol 1,4,5-trisphosphate and diacylglycerol allows comprehensive analysis of phospholipase C-initiated signalling in single neurons.Authors: Nelson Et al.J Neurochem2008;107:602
- IGF-I inhibition of apoptosis is associated with decreased expression of prostate apoptosis response-4.Authors: Chung Et al.J Endocrinol2007;194:77
- Protein kinase C dependent inhibition of the heteromeric Kir4.1-Kir5.1 channel.Authors: Rojas Et al.Biochim Biophys Acta2007;1768:2030
- PI-3-kinase-dependent membrane recruitment of centaurin-α2 is essential for its effect on ARF6-mediated actin cytoskeleton reorganisation.Authors: Venkateswarlu Et al.J Neurosci2007;120:792
- GSK-3beta acts downstream of PP2A and the PI 3-kinase-Akt pathway, and upstream of caspase-2 in ceramide-induced mitochondrial apoptosis.Authors: Lin Et al.J Cell Sci2007;120:2935
- Sphingosine 1-phosphate induces endothelial nitric-oxide synthase activation through phosphorylation in human corpus cavernosum.Authors: Bianca Et al.J Cell Biochem2006;316:703
- Regulation of nuclear factor κB in the hippocampus by group I metabotropic glutamate receptors.Authors: O'Riordan Et al.J Neurosci2006;26:4870
- Differential reelin-induced enhancement of NMDA and AMPA receptor activity in the adult hippocampus.Authors: Qiu Et al.Mol Cancer Ther2006;26:12943
- Myosin light chain kinase is not a regulator of synaptic vesicle trafficking during repetitive exocytosis in cultured hippocampal neurons.Authors: Tokuoka and GodaJ Neurosci2006;26:11606
- LY 294002 inhibits adenosine receptor activation by a mechanism independent of effects on PI-3 kinase or casein kinase II.Authors: Searl and SilinskyPurinergic Signal2005;1:389
- Mechanisms underlying the relaxation response induced by bradykinin in the epithelium-intact guinea-pig trachea in vitro.Authors: Schlemper Et al.Br J Pharmacol2005;145:740
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10nM Wortmannin
Treatment of fibroblasts for 4 hours prior to staining for LC3 and P62.
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天然产物,大多都有颜色,
存在干扰,多数情况下需要做样品的阴性对照,
尽量能用荧光的方法,
之前我们做过,将两个试剂盒的方法合并后,做的,
效果还可以
2.微孔板:选择实验所需的板条数。剩余不用的部分随同干燥剂密封放回原袋。 1.包被有抗抑制素bB亚单位抗体的微孔板(Anti-InhibinB-CoatedMicrotitrationStrips):96孔,2-8ºC下密封保存。
2.标准品A/样本稀释液(InhibinBStandardA/SampleDiluent):2ml×1瓶,胎牛血清,含0pg/mL二聚体抑制素B。使用前2-8ºC下保存。打开后2-8ºC下可保存2周。于-20ºC可长期保存。
3.标准品B-G(InhibinBStandardsB-G):1ml×6瓶,胎牛血清,含二聚体抑制素B的浓度为10,30,100,250,500,1000pg/mL。使用前2-8ºC下保存。打开后2-8ºC下可保存2周。于-20ºC可长期保存。
4.质控(InhibinBControls):1ml×2瓶,浓度I、II,胎牛血清,含低浓度和高浓度的抑制素A二聚体。使用前2-8ºC下保存。打开后2-8ºC下可保存2周。于-20ºC可长期保存。
5.样本稀释液A(InhibinBSampleBufferA):10ml×1瓶,2-8ºC下保存。
6.样本稀释液B(InhibinBSampleBufferB):10ml×1瓶,2-8ºC下保存。
7.抗体-生物素结合物(InhibinBAntibody-BiotinConjugate)(即用型):10ml×1瓶,含生物素标记的抗抑制素a亚单位抗体。2-8ºC下保存。
8.酶结合物(浓缩)(Streptavidin-EnzymeConjugate)(即用型):10ml×1瓶,含链霉和素-辣根过氧化物酶结合物。2-8ºC下保存。
9.TMB底物溶液(TMBChromogenSolution):15ml×1瓶,2-8ºC下保存。
10.终止液(StoppingSolution):15ml×1瓶,为0.2M的硫酸。2-8ºC下保存。
11.浓缩洗液(WashConcentrate):100ml×1瓶,2-8ºC下保存。 操作前所有试剂都应平衡至室温(~25ºC)并充分混匀。标准品、质控及待测样本需同时做双份。
1.取出实验所需板条,并记录各孔位置。
2.在对应的孔中加入标准品、质控及待测样本各50ul。
3.每孔加入25ul样本稀释液A。
4.每孔加入25ul样本稀释液B。
5.封板,以300-400rpm速度震荡,室温下过夜(14-18小时)。
6.洗板3次,在吸水纸上拍干。
7.每孔加入50ul抗体-生物素结合物。
8.封板,以500-700rpm速度震荡,室温下孵育1.5小时。
9.洗板6次,在吸水纸上拍干。
10.每孔加入50ul链霉和素-辣根过氧化物酶结合物。
11.封板,以500-700rpm速度震荡,室温下孵育20分钟。
12.洗板6次,洗完最后一次以后,将洗液在孔内停留15分钟再除去。在吸水纸上拍干。
13.在每孔中加入100ulTMB底物溶液。
14.以500-700rpm速度震荡,室温下避光孵育15-30分钟。
15.每孔加入100ul终止液。
16.30分钟内在450nm处读数。
注:必须以零标准作空白。如果能做到双波长测定,可在600或620nm处读数,将600(620)nm吸收值从450nm处减去,这样可以减少光学误差。 使用血清样本,静脉穿刺术采集血液。样本于2-8ºC可保存24小时,-20ºC或以下可保存30天。避免反复冻融样本。不应采用溶血或脂血的样本。冰冻的样本在实验前应解冻,并充分混匀。向左转|向右转
RMgX + RX = R-R + MgX2.
这个反应需要的能量比生成格氏试剂的高,
因此降低反应温度是第一个选择。
其次, 增加镁得摩尔比, 让 RX与镁有更多机会反应, 而不是与RMgX。
第三, 降低RX的浓度, 即用更多的溶剂, 因为溶剂和格氏试剂有很显著的溶剂络合。
第四, 缓慢滴加RX., 即降低RX.在反应体系的浓度。
第五, 增加搅拌速率, 即, 让RX.与镁有更好的接触。

