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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我用conA和LPS分别刺激脾细胞,当然细胞均有明显的增殖,可是如果把这两种有丝分裂原同时用来刺激细胞时,细胞的增殖明显受到了抑制,比起单独采用ConA或者LPS都低很多。conA和LPS分别刺激T细胞和B细胞的增殖,两者同时加入时,作用应该更强,为什么反而大幅减弱呢?
此外,我研究的这种成分单独刺激皮细胞时,细胞也有一定程度的增殖,但是如果这种物质与conA联合使用时,比起单独用conA时,细胞的增殖程度明显降低了一些,这又是什么原因呢?难道可以解释为我研究的这种物质有类似于LPS的作用——刺激B细胞增殖?
如果真的是这样来解释,哪位大侠能把第一个现象(conA和LPS共同作用产生拮抗)的机理帮我解释一下呢?
当抗体结合到示踪剂上时,340nm的激发光激发铕标分子,导致能量转移到Alexa Fluor 647染料上,结果产生665nm的发射光。荧光的强度与样品中的cAMP含量成反比。
本试剂盒用于检测在GPCR激动剂刺激下活细胞或者细胞膜制备品产生的cAMP。对于偶联Gαs的受体,激动剂刺激导致665nm的荧光强度降低,而拮抗剂则可以逆转这一效应;对于偶联Gαi的受体,在激动剂刺激的同时用forskolin刺激cAMP产生,那么激动剂则抑制forskolin诱导的cAMP的生成,因此对照只给forskolin的细胞组可以通过665nm荧光强度的增加反应激动剂的效应。
该试剂盒的灵敏度很高,室温下反应在20h内是稳定的。本试剂盒适用于在384孔板中进行24μl的微量分析。
2.保存条件
避光2~4℃保存,过期时间见装。
3.盒内试剂
cAMP标准品:1管,1ml。(50μM)
生物素标记的cAMP(b-cAMP):1管,25μl。
铕标的抗生物素蛋白链菌素:1管,25μl。
荧光标记的cAMP抗体:1管,40μl。
检测缓冲液:1瓶,25ml。
4.需要自配的其他溶液
l Hank’s balanced salt solution (HBSS): NaCl 8.0g、CaCl2 0.14g、KCl 0.4g、 KH2PO4 0.06g、Na2HPO4?7H2O0.09g、MgCl2.6H2O0.10 g、MgSO4.7H2O0.10 g、NaHCO30.35g、葡萄糖1.0g,加H2O至 1000ml (用7.5%NaHCO调节PH值=7.4)
l Versene消化液(1L):EDTA 0.372 g,NaCl 8.0g,KCl 0.20 g,KH2PO40.20g,Na2HPO4 1.15 g,D-glucouse 0.2 g,pH 7.4
l HEPES缓冲液(1mol/L):取2.383gHEPES溶于10ml去离子水中。
l 7.5%BSA溶液:取0.75gBSA溶于10ml去离子水中
l 0.5M IBMX溶液:11.11mg IBMX溶于100μl DMSO中,-20℃冻存。
l 刺激缓冲液(SB):14 ml HBSS(1×)+75μlHEPES(1mol/L)+200μlBSA (7.5%)。(注:在测定细胞cAMP时,反应缓冲液中要加入IBMX 0.5mmol/L)
l 吗啡贮存液(10mM):盐酸吗啡37.585mg溶于10ml生理盐水中,0.22μm滤膜过滤除菌,4℃保存备用。
l 纳络酮母液(100mM):纳络酮4mg溶于100μl 生理盐水中,用时工作液按照1:500稀释,溶剂为含有IBMX的反应缓冲液。
抑制剂刺激细胞后,需要用PBS清洗后再做后续实验吗
我做的是细胞因子的刺激和抑制某条通路后观察是否有影响,分组为空白组,空白+抑制剂,刺激组,刺激+抑制剂,最开始用的单因素方差分析,LSD-T和SNK-Q检验,但是同学说我这里面有两个处理因素,所以不能单因素方差分析,应该直接空白和空白+抑制,空白和刺激,刺激和刺激+抑制剂进行独立样本T检验,现在脑子是混乱的,拜托园子里的大神们帮我看看,感激不尽!!

