Biological Activity
Potent inhibitor of sarco-endoplasmic reticulum Ca2+-ATPases. Causes ER stress; can be used to induce autophagy in mammalian cells.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
- Differential effects of endoplasmic reticulum stress-induced autophagy on cell survival.Ding et al.J.Biol.Chem., 2007;282:4702
- Kinetics of thapsigargin-Ca2+-ATPase (sarcoplasmic reticulum) interaction reveals a two-step binding mechanism and picomolar inhibition.Davidson and VarholJ.Biol.Chem., 1995;270:11731
- A tool coming of age: thapsigargin as an inhibitor of sarco-endoplasmic reticulum Ca2+-ATPases.Treiman et al.TiPS, 1998;19:131
- Specific substitutions at amino acid 256 of the sarcoplasmic/endoplasmic reticulum Ca2+ transport ATPase mediate resistance to thapsigargin in thapsigargin-resistant hamster cells.Yu et al.J.Biol.Chem., 1998;273:3542
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Citations for Thapsigargin
The citations listed below are publications that use Tocris products.Selected citations for Thapsigargin include:
41Citations: Showing 1 - 10
- Swelling-activated Ca2+ channels trigger Ca2+ signals in Merkel cells.Authors: Haeberle Et al.MBio;3:e1750
- Caspase-mediated cleavage of IRE1 controls apoptotic cell commitment during endoplasmic reticulum stress.Authors: Shemorry Et al.Elife2019;8
- Dysfunctional autophagy following exposure to pro-inflammatory cytokines contributes to pancreatic β-cell apoptosis.Authors: Lambelet Et al.Cell Death Dis2018;9:96
- Hyperactivation of nuclear receptor coactivators induces PERK-dependent cell death.Authors: Hossain Et al.Oncotarget2018;9:11707
- Mechanical regulation of stem-cell differentiation by the stretch-activated Piezo channel.Authors: He Et al.Nature2018;555:103
- IP3 Receptors Preferentially Associate with ER-Lysosome Contact Sites and Selectively Deliver Ca2+ to Lysosomes.Authors: Atakpa Et al.Cell Rep2018;25:3180
- Mitochondria-Derived Vesicles Deliver Antimicrobial Reactive Oxygen Species to Control Phagosome-Localized Staphylococcus aureus.Authors: Abuaita Et al.Cell Host Microbe2018;24:625
- Increased sensitivity to apoptosis upon endoplasmic reticulum stress-induced activation of the unfolded protein response in chemotherapy-resistant malignant pleural mesothelioma.Authors: Xu Et al.Br J Cancer2018;119:65
- Calcium-regulatory proteins as modulators of chemotherapy in human neuroblastoma.Authors: Florea Et al.Oncotarget2017;8:22876
- Role of two sequence motifs of mesencephalic astrocyte-derived neurotrophic factor in its survival-promoting activity.Authors: Mätlik Et al.Cell Death Dis2016;6:e2032
- NCOA3 coactivator is a transcriptional target of XBP1 and regulates PERK-eIF2α-ATF4 signalling in breast cancer.Authors: Gupta Et al.Oncogene2016;35:5860
- Presynaptic Spike Timing-Dependent Long-Term Depression in the Mouse Hippocampus.Authors: Andrade-Talavera Et al.Cereb Cortex2016;26:3637
- Control of sensory neuron excitability by serotonin involves 5HT2C receptors and Ca2+-activated chloride channels.Authors: Salzer Et al.Neuropharmacology2016;110 (A):277
- α-1 adrenoreceptors modulate GABA release onto ventral tegmental area DA neurons.Authors: Velásquez-Martínez Et al.Pflugers Arch2015;88:110
- Binding of αherpesvirus Glycoprotein H to Surface α4β1-Integrins Activates Calcium-Signaling Pathways and Induces Phosphatidylserine Exposure on the Plasma Membrane.Authors: Azab Et al.Endocrinology2015;6
- Calcium release through P2X4 activates calmodulin to promote endolysosomal membrane fusion.Authors: Cao Et al.PLoS One2015;209:879
- Differential regulation of GnRH secretion in the preoptic area (POA) and the median eminence (ME) in male mice.Authors: Glanowska and MoenterNeuropharmacology2015;156:231
- TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis.Authors: Shambharkar Et al.J Cell Biol2015;10:e0127480
- Caffeine Modulates Vesicle Release and Recovery at Cerebellar Parallel Fibre Terminals, Independently of Calcium and Cyclic AMP Signalling.Authors: Dobson Et al.J Neuroinflammation2015;10:e0125974
- Orai1 and STIM1 mediate SOCE and contribute to apoptotic resistance of pancreatic adenocarcinoma.Authors: Kondratska Et al.PLoS One2014;1843:2263
- Downregulation of BRCA1 protein in BCR-ABL1 leukemia cells depends on stress-triggered TIAR-mediated suppression of translation.Authors: Podszywalow-Bartnicka Et al.Biochim Biophys Acta2014;13:3727
- Activation of ERK1/2 by store-operated calcium entry in rat parotid acinar cells.Authors: Soltoff and LannonPLoS One2013;8:e72881
- Optogenetic probing and manipulation of the calyx-type presynaptic terminal in the embryonic chick ciliary ganglion.Authors: Egawa Et al.PLoS One2013;8:e59179
- Endoplasmic reticulum membrane reorganization is regulated by ionic homeostasis.Authors: Varadarajan Et al.PLoS One2013;8:e56603
- Activation of group I metabotropic glutamate receptors potentiates heteromeric kainate receptors.Authors: Rojas Et al.Mol Pharmacol2013;83:106
- Recruitment of the intracellular Ca2+ by ultrashort electric stimuli: the impact of pulse duration.Authors: Semenov Et al.Cell Calcium2013;54:145
- The expression and relaxant effect of bitter taste receptors in human bronchi.Authors: Grassin-Delyle Et al.Respir Res2013;14:134
- Bradykinin-induced Ca2+ signaling in human subcutaneous fibroblasts involves ATP release via hemichannels leading to P2Y12 receptors activation.Authors: Pinheiro Et al.Cell Commun Signal2013;11:70
- Asynchronous Ca2+ current conducted by voltage-gated Ca2+ (CaV)-2.1 and CaV2.2 channels and its implications for asynchronous neurotransmitter release.Authors: Few Et al.Proc Natl Acad Sci U S A2012;109:E452
- ER stress induces anabolic resistance in muscle cells through PKB-induced blockade of mTORC1.Authors: Deldicque Et al.PLoS One2011;6:e20993
- Associative, bidirectional changes in neural signaling utilizing NMDA receptor- and endocannabinoid-dependent mechanisms.Authors: Li and BurrellLearn Mem2011;18:545
- β1-Adrenergic receptors activate two distinct signaling pathways in striatal neurons.Authors: Meitzen Et al.J Neurochem2011;116:984
- Dysregulation of presynaptic calcium and synaptic plasticity in a mouse model of 22q11 deletion syndrome.Authors: Earls Et al.PLoS One2010;30:15843
- Plasticity of burst firing induced by synergistic activation of metabotropic glutamate and acetylcholine receptors.Authors: Moore Et al.Neuron2009;61:287
- Calcium homeostasis and cone signaling are regulated by interactions between calcium stores and plasma membrane ion channels.Authors: Szikra Et al.PLoS One2009;4:e6723
- H(2)O(2)-mediated modulation of cytosolic signaling and organelle function in rat hippocampus.Authors: Gerich Et al.J Neurosci2009;458:937
- A crucial role for cAMP and protein kinase A in D1 DA receptor regulated intracellular calcium transients.Authors: Dai Et al.Neurosignals2008;16:112
- The role of protein kinase A in the ethanol-induced increase in spontaneous GABA release onto cerebellar Purkinje neurons.Authors: Kelm Et al.J Neurophysiol2008;100:3417
- Calcium release from presynaptic internal stores is required for ethanol to increase spontaneousγ-aminobutyric acid release onto cerebellum Purkinje neurons.Authors: Kelm Et al.Cell Cycle2007;323:356
- Mechanism of graded persistent cellular activity of entorhinal cortex layer v neurons.Authors: Fransen Et al.Neuron2006;49:735
- Two coincidence detectors for spike timing-dependent plasticity in somatosensory cortex.Authors: Bender Et al.J Neurosci2006;26:4166
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Thapsigargin was used to block SERCA in activated CD8+ T-cells. This maintained high intracellular Ca2+, leading to increased activation of CD8+ T-cells as measured by IFNg production via intracellular flow cytometry.
This compound is often used to deplete intracellular calcium stores in cell culture or other applications. It worked as expected in our tissue preparations.
Cytosolic Ca2+ sensitive indicator, Fura-2 was loaded in RBL-2H3 cells and excited alternately at 340 and 380 nm and fluorescence was captured at 510 nm.ER store was depleted with 500 nM thapsigargin which promotes store-operated Ca2+ entry via STIM1/Orai1 pathway into the cells upon addition of 2 mM extracellular Ca2+.RBL-2H3 cells were loaded with FURA-2AM (2 μM) and Pluronic F127 (0.02%) for 30 min followed by washing with normal-Tyrode solution. Cells were stimulated at 1 Hz frequency and fluorescence intensities were measured.
Used it as a positive control to test if the mechanism of a novel small molecule (compound x) was to reduce ER stress.Cells were treated with 5um Thapsigargin for 24 hours, and splicing of XBP1 was determined by semi-quantitative RT-PCR
for calcium release from the ER
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2.SNaPshot 该技术由美应用物公司(ABI)发基于荧光标记单碱基延伸原理型技术称测序主要针等通量SNP型项目含测序酶、四种荧光标记ddNTP、紧临态位点5’-端同度延伸引物PCR产物模板反应体系引物延伸碱基即终止经ABI测序仪检测根据峰移位置确定该延伸产物应SNP位点根据峰颜色知掺入碱基种类确定该本基型于PCR产物模板通重PCR反应体系获通用于10-30SNP位点析
期待着高手们的指点,谢谢。
不是很相信网上的商家,遂求助各位路过的大虾们,有用过的吗?这些酶特异性怎样啊?贵不贵,跟我直接测序相比,有优势吗?
根据发展历史、影响力、测序原理和技术不同等,主要有以下几种:大规模平行签名测序(Massively Parallel Signature Sequencing, MPSS)、聚合酶克隆(Polony Sequencing)、454焦磷酸测序(454 pyrosequencing)、Illumina (Solexa) sequencing、ABI SOLiD sequencing、离子半导体测序(Ion semiconductor sequencing)、DNA 纳米球测序 (DNA nanoball sequencing)等。 基因分析仪(即DNA测序仪),采用毛细管电泳技术取代传统的聚丙烯酰胺平板电泳,应用该公司专利的四色荧光染料标记的ddNTP(标记终止物法),因此通过单引物PCR测序反应,生成的PCR产物则是相差1个碱基的3'末端为4种不同荧光染料的单链DNA混合物,使得四种荧光染料的测序PCR产物可在一根毛细管内电泳,从而避免了泳道间迁移率差异的影响,大大提高了测序的精确度。由于分子大小不同,在毛细管电泳中的迁移率也不同,当其通过毛细管读数窗口段时,激光检测器窗口中的CCD(charge-coupled device)摄影机检测器就可对荧光分子逐个进行检测,激发的荧光经光栅分光,以区分代表不同碱基信息的不同颜色的荧光,并在CCD摄影机上同步成像,分析软件可自动将不同荧光转变为DNA序列,从而达到DNA测序的目的。分析结果能以凝胶电泳图谱、荧光吸收峰图或碱基排列顺序等多种形式输出。
【之前构建的4个重组质粒里也有两个出现这样的问题(aagtcc变成agtcc),但是酶切能切出插入的片段】
目前问测序公司得到的答复是样品可能不是单克隆,测序结果是不准确的,建议重新挑取单克隆测序。
不知道还有没有别的方法?
以这些标准看,目前的基因组测序结果,还没有一个是完美的。
人类基因组:缺点在哪里?
首先,人类基因组还不够精确。人是“二倍体”,也就是有一半遗传物质来自父亲,一半遗传物质来自母亲,且在受精卵形成过程中,还会发生基因重组,这是人类遗传多样性的来源之一。科学家们需要更精确的“单倍型”数据,这样基因组才够“完美”,而这种“完美”正是研究者们追求的目标。
其次,人类基因组还不够多元。
按照传统的人种分类,人类按照肤色黑白黄棕,被粗分为四大类:尼格罗人种、高加索人种、蒙古人种、澳大利亚人种。基因组测序数据是从高加索人种开始的,人类基因组计划是人类的标准参考基因组,也是高加索人种的标准参考基因组。文特尔的基因组,测序对象是他自己,同样是高加索人种。
然而,从基因组研究的角度,为了尽可能地包括各种遗传背景,需要为更多族裔建立自己的参考基因组。
资料来源:http://www.mv163.cn/jkgl/news/2015/1224/5227.html
目的片段用kpn1和xho1双酶切,然后连接到PGL4.10上,结果用RPV3测序出来目的序列是反向互补的,这是什么原因导致的?
你所说的“测序酶”在一般情况下就是“聚合酶”。
因为目前的的测序方法主要就是借助聚合反应。
当然,有些测序方法(比如曾经的SOLiD系统)是利用连接反应,那么它用的“测序酶”肯定是连接酶。
估计,也就是这个原因有人把它。

