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Boston Biochem/Thapsigargin/1138/1
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Boston Biochem
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1138/1
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Product Details
Citations (41)
Reviews (6)
Biological Activity Technical DataBackground Product DatasheetsCalculators

Biological Activity

Potent inhibitor of sarco-endoplasmic reticulum Ca2+-ATPases. Causes ER stress; can be used to induce autophagy in mammalian cells.

Technical Data

M.Wt:
650.76
Formula:
C34H50O12
Solubility:
Soluble to 100 mM in DMSO
Purity:
≥97%
Storage:
Desiccate at -20°C
CAS No:
67526-95-8

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

  1. Differential effects of endoplasmic reticulum stress-induced autophagy on cell survival.Ding et al.J.Biol.Chem., 2007;282:4702
  2. 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
  3. A tool coming of age: thapsigargin as an inhibitor of sarco-endoplasmic reticulum Ca2+-ATPases.Treiman et al.TiPS, 1998;19:131
  4. 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

Product Datasheets

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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

  1. Swelling-activated Ca2+ channels trigger Ca2+ signals in Merkel cells.Authors: Haeberle Et al.MBio;3:e1750
  2. Caspase-mediated cleavage of IRE1 controls apoptotic cell commitment during endoplasmic reticulum stress.Authors: Shemorry Et al.Elife2019;8
  3. Dysfunctional autophagy following exposure to pro-inflammatory cytokines contributes to pancreatic β-cell apoptosis.Authors: Lambelet Et al.Cell Death Dis2018;9:96
  4. Hyperactivation of nuclear receptor coactivators induces PERK-dependent cell death.Authors: Hossain Et al.Oncotarget2018;9:11707
  5. Mechanical regulation of stem-cell differentiation by the stretch-activated Piezo channel.Authors: He Et al.Nature2018;555:103
  6. IP3 Receptors Preferentially Associate with ER-Lysosome Contact Sites and Selectively Deliver Ca2+ to Lysosomes.Authors: Atakpa Et al.Cell Rep2018;25:3180
  7. Mitochondria-Derived Vesicles Deliver Antimicrobial Reactive Oxygen Species to Control Phagosome-Localized Staphylococcus aureus.Authors: Abuaita Et al.Cell Host Microbe2018;24:625
  8. 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
  9. Calcium-regulatory proteins as modulators of chemotherapy in human neuroblastoma.Authors: Florea Et al.Oncotarget2017;8:22876
  10. 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
  11. NCOA3 coactivator is a transcriptional target of XBP1 and regulates PERK-eIF2α-ATF4 signalling in breast cancer.Authors: Gupta Et al.Oncogene2016;35:5860
  12. Presynaptic Spike Timing-Dependent Long-Term Depression in the Mouse Hippocampus.Authors: Andrade-Talavera Et al.Cereb Cortex2016;26:3637
  13. Control of sensory neuron excitability by serotonin involves 5HT2C receptors and Ca2+-activated chloride channels.Authors: Salzer Et al.Neuropharmacology2016;110 (A):277
  14. α-1 adrenoreceptors modulate GABA release onto ventral tegmental area DA neurons.Authors: Velásquez-Martínez Et al.Pflugers Arch2015;88:110
  15. 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
  16. Calcium release through P2X4 activates calmodulin to promote endolysosomal membrane fusion.Authors: Cao Et al.PLoS One2015;209:879
  17. Differential regulation of GnRH secretion in the preoptic area (POA) and the median eminence (ME) in male mice.Authors: Glanowska and MoenterNeuropharmacology2015;156:231
  18. TMEM203 Is a Novel Regulator of Intracellular Calcium Homeostasis and Is Required for Spermatogenesis.Authors: Shambharkar Et al.J Cell Biol2015;10:e0127480
  19. 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
  20. Orai1 and STIM1 mediate SOCE and contribute to apoptotic resistance of pancreatic adenocarcinoma.Authors: Kondratska Et al.PLoS One2014;1843:2263
  21. 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
  22. Activation of ERK1/2 by store-operated calcium entry in rat parotid acinar cells.Authors: Soltoff and LannonPLoS One2013;8:e72881
  23. Optogenetic probing and manipulation of the calyx-type presynaptic terminal in the embryonic chick ciliary ganglion.Authors: Egawa Et al.PLoS One2013;8:e59179
  24. Endoplasmic reticulum membrane reorganization is regulated by ionic homeostasis.Authors: Varadarajan Et al.PLoS One2013;8:e56603
  25. Activation of group I metabotropic glutamate receptors potentiates heteromeric kainate receptors.Authors: Rojas Et al.Mol Pharmacol2013;83:106
  26. Recruitment of the intracellular Ca2+ by ultrashort electric stimuli: the impact of pulse duration.Authors: Semenov Et al.Cell Calcium2013;54:145
  27. The expression and relaxant effect of bitter taste receptors in human bronchi.Authors: Grassin-Delyle Et al.Respir Res2013;14:134
  28. 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
  29. 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
  30. ER stress induces anabolic resistance in muscle cells through PKB-induced blockade of mTORC1.Authors: Deldicque Et al.PLoS One2011;6:e20993
  31. Associative, bidirectional changes in neural signaling utilizing NMDA receptor- and endocannabinoid-dependent mechanisms.Authors: Li and BurrellLearn Mem2011;18:545
  32. β1-Adrenergic receptors activate two distinct signaling pathways in striatal neurons.Authors: Meitzen Et al.J Neurochem2011;116:984
  33. Dysregulation of presynaptic calcium and synaptic plasticity in a mouse model of 22q11 deletion syndrome.Authors: Earls Et al.PLoS One2010;30:15843
  34. Plasticity of burst firing induced by synergistic activation of metabotropic glutamate and acetylcholine receptors.Authors: Moore Et al.Neuron2009;61:287
  35. 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
  36. H(2)O(2)-mediated modulation of cytosolic signaling and organelle function in rat hippocampus.Authors: Gerich Et al.J Neurosci2009;458:937
  37. A crucial role for cAMP and protein kinase A in D1 DA receptor regulated intracellular calcium transients.Authors: Dai Et al.Neurosignals2008;16:112
  38. 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
  39. 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
  40. Mechanism of graded persistent cellular activity of entorhinal cortex layer v neurons.Authors: Fransen Et al.Neuron2006;49:735
  41. Two coincidence detectors for spike timing-dependent plasticity in somatosensory cortex.Authors: Bender Et al.J Neurosci2006;26:4166

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Reviews for Thapsigargin

Average Rating: 4.3(Based on 6 Reviews)

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Usage Information
By Anonymouson 06/20/2020
Application: Species:Human

100 nM

PMID: 31453810Reference

Thapsigargin Blocks Re-Uptake of Ca2+ Into ER leading to increased T-cell activation by IFNg
By Li Zhuon 01/23/2019
Application: Species:Human

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.


Intracellular calcium depletion
By Anonymouson 11/19/2018
Application: Species:Mouse

This compound is often used to deplete intracellular calcium stores in cell culture or other applications. It worked as expected in our tissue preparations.


Thapsigargin elicits store-operated Ca2+ entry after ER store depletion in RBL-2H3 cells.
By Niroj Shresthaon 10/27/2018
Application: Species:Rat

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.


XBP1 splicing
By Preethi Vijayarajon 05/01/2018
Application: Species:Human

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


Thapsigargin calcium assay
By Anonymouson 12/12/2017
Application: Species:Human

for calcium release from the ER


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PCR程延伸阶段温度升72摄氏度左右高温度所溶液四种脱氧核苷酸需要耐高温Taq DNA聚合酶作用据碱基互补配原则合新DNA链
正在做bmp4启动子的一段序列,两端分别加的Sac1和Nhe1的位点,pcr之后跑胶显示条带位置正确,回收后分别与pgemt-easy载体和peasy-t5zero载体连接,连接之后单酶切和双酶切均显示连接成功,但测序结果就是不对,已经测了3次,求哪位大神指教一二,多谢了!!难道真的是pcr出的序列本来就是错的?而且连接peasy-t5zero载体后双酶切切不开。我用的Sac1和Nhe1是Thermo的,正在尝试换takara的重新双酶切。
pgem-teasy载体插入片段位点两侧各有一个EcoR1的位点。

1.TaqMan探针 针染色体同SNP位点别设计PCR引物TaqMan探针进行实荧光PCR扩增探针5’-端3’-端别标记报告荧光基团淬灭荧光基团溶液存PCR产物该探针与模板退火即产适合于核酸外切酶性底物探针5’-端连接荧光探针切割破坏两荧光间PRET发荧光通用于少量SNP位点析
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
我的实验中,为了检测一个明确报道的多态性位点(该位点为酶切位点)在细胞中的多态性状态,能不能用高保真酶扩增含该位点的一段序列后,直接测序?还是应该先做酶切,在测序?测序是需要做一个t载体克隆不?

期待着高手们的指点,谢谢。
我在做基因敲除实验(CRISPR/Cas9),实验室一直都是直接送测序验证的,又看到资料说,有用T7E1酶的,也有用SURVEYOR的。
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?
高通量测序技术(High-throughput sequencing)又称“下一代”测序技术(Next-generation sequencing technology),以能一次并行对几十万到几百万条DNA分子进行序列测定和一般读长较短等为标志。
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。
片段长度143bp,酶切位点kpn1和Hind3,连入pGL3b载体,测序引物公司用的pGL3-,第一次反向测序不成功,原因是质粒出现重叠峰,正向加测后成功,对比发现下游引物的酶切位点(Hind3)少了一个碱基(aagtcc变成aagct),重组质粒酶切切不出插入片段;之后将测序成功的质粒重新转化DH5α,再提质粒酶切还是切不出来。质粒小提试剂盒Qiagen的,提取菌液1.5mL,浓度608.2ng/uL,酶切质粒2ug。
【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?
在基因组研究领域,人们对数据的可信度有一个基本的要求:单个碱基越准确越好,对单个碱基的覆盖深度越多倍越好,对整个基因组测得越完整越好,测序的“缺口(Gap)”越少越好。

  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?

我的目的蛋白D2,是膜表达蛋白,构建了三种载体的重组质粒,分别为CMV,pCDNA3.1,pEGFP-N1,三个重组质粒酶切正确、测序正确,也曾表达过,可以砸出相应大小的目的条带。大提过后,有近一个半月没有进行转染表达,突然发现我的质粒不能砸出目的条带了,甚至外源性的anti-Flag都砸不到(pEGFP重组质粒转染可以观察到微弱荧光,比以前较少),已经重修小提过,也重新进行了酶切,进行测序,都没有问题,现在就是砸不到条带(阳性对照的Flag每次都可以),请各位大神帮忙!!!!!!!!!!!!!!!!!!!!

没有专门的“测序酶”的说法。
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。
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