Description:
SMAD2/3ResponsiveLuciferaseReporterNIH/3T3StableCellLineisderivedfrommousefibroblast,andstablyexpressfireflyluciferasereportergeneunderthecontrolofSMAD2/3responseelement. Thiscelllineisanidealcellularmodelformonitoringtheactivation ofTGFbeta SignalingPathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.
Principle:
Smadproteinsaretranscriptionfactorsthatrespondtotransforminggrowthfactor-β(TGFβ)signaling,whereTGFβinducesitsmembranereceptorstodirectlyactivateSmadproteins. TheseactivatedSmadscomplexwithSmad4(co-Smad),translocatefromcytoplasmintonucleusandbindtotargetpromoterregiontoregulategenetranscriptions. DysfunctioninTGFβpathwayleadstoimmunosuppressionandangiogenesis,whichcanmakecancermoreinvasive.
SignosishasdevelopedSMAD/TGFbluciferasereporterstablecelllinebyco-transfectingSMADluciferasereportervectorandhygromycinexpressionvector. ThehygromycinresistantclonesweresubsequentlyscreenedforTGFb1-inducedluciferaseactivity. ThecelllinecanbeusedasareportersystemformonitoringtheactivationofSMADtriggeredbystimulitreatment,suchasTGFb1,andgeneoverexpressionandgeneknockdown.
![](images/signosisinc/201709/luciferase reporter principle diagram v2.png)
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
![](http://www.signosisinc.com/data:image/png;base64,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)
AnalysisofSMAD/TGFbetaLuciferaseReporterNIH/3T3StableCellLine.NIH/3T3cellsstablyexpressingSMADluciferasereporterweretreatedwith20ng/mLTGF-beta1inDMEM+0.1%FBSfor16hours. TGF-betaisoformsinducedabout8-foldincreaseinSMAD-luciferasereporteractivity.
蚂蚁淘电商平台
ebiomall.com
公司简介
蚂蚁淘(www.ebiomall.cn)是中国大陆目前唯一的生物医疗科研用品B2B跨境交易平台,
该平台由多位经验丰富的生物人和IT人负责运营。蚂蚁淘B2B模式是指客户有采购意向后在蚂蚁
淘搜索全球供应信息,找到合适的产品后在蚂蚁淘下单,然后蚂蚁淘的海外买手进行跨境采购、
运输到中国口岸,最后由蚂蚁淘国内团队报关运输给客户...
正品保证: 全球直采 在线追溯
蚂蚁淘所有产品都是自运营的,我们已经跟国外多家厂方建立品牌推广合作关系, 获得对方的支持和授权; 同时客户可以通过订单详情查看到货物从厂方至客户的所有流程, 确保货物的来源; 正规报关,提供13%增值税发票。
及时交付: 限时必达 畅选无忧
蚂蚁淘的运营团队都是有着多年经验的成员,他们熟悉海外采购、仓储物流、报关等环节; 同时通过在线的流程监控,蚂蚁淘的进口速度比传统企业提高了50%以上, 部分产品甚至能做到7-10天到货,即蚂蚁淘的“时必达”服务。
轻松采购: 在线下单 简单省事
蚂蚁淘的价格是真实透明的,并且具有很大的价格优势,不需要繁杂的询价比价; 报价单与合同可以直接在线生成或打印;就像在京东购物一样, 您的鼠标点击几 次即完成在蚂蚁淘的采购,订单详情会告诉您所有进程。
售后申请: 耐心讲解 优质服务
蚂蚁淘提供的产品在使用过程中如因产品质量问题有售后需求时, 您可通过我的订单提交您的“申请售后”, 蚂蚁淘产品顾问会第一时间为您处理, 在售后服务过程中如遇到问题也可致电蚂蚁淘客服热线:4000-520-616。
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For amplification of cognate sequences from different organisms, or for "evolutionary PCR", one may increase the chances of getting product by designing
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稳定表达细胞株(稳转细胞株)指在细胞中持续稳定表达特定基因或干扰特定基因表达。稳转细胞株的目的基因质粒DNA整合到细胞染色体上,使细胞长期稳定表达该基因。 在基因功能的
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江苏吉锐生物技术有限公司在发布的稳转细胞株构建(过表达、沉默)--稳定遗传,优于慢病毒供应信息,浏览与稳转细胞株构建(过表达、沉默)--稳定遗传,优于慢病毒相关的产品或在搜索更多与稳转细胞株构建(过表达、沉默)--稳定遗传,优于慢病毒相关的内容。
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稳定转染和瞬时转染,细胞转染的步骤基本相同,只是在转染后的细胞处理会不一样。由于瞬时转染只能维持几天,故在转染12~72后就可以在蛋白或基因水平进行相应的检测了。稳转细胞系通常需要通过一些选择性标记反复筛选,得到稳定转染的同源细胞系。通常构建的载体上有两个抗性基因,如,氨苄青霉素和新霉素。那么建立稳转细胞系时该用哪种抗生素来筛细胞呢?很多真核表达质粒上都有两个抗性基因,但一般其中只有一个抗性可以用来筛细胞。其中氨苄青霉素对细胞是没有筛选...
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北京英茂盛业生物科技有限公司在发布的MDCK-EGFP稳定细胞株(犬肾细胞EGFP稳转细胞系)供应信息,浏览与MDCK-EGFP稳定细胞株(犬肾细胞EGFP稳转细胞系)相关的产品或在搜索更多与MDCK-EGFP稳定细胞株(犬肾细胞EGFP稳转细胞系)相关的内容。
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PREPARING CELLSBring up HUVEC and fibroblasts in M199/10% FBS/Pen-Strep (1:100) 1-2 days before beading. Switch medium to EGM-2 (Clonetics) the day before bead
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hl-60细胞是一种用于实验室研究某些血细胞如何形成的细胞株。在培养基中补充牛胎儿血清.HEPES,抗生素类化学物质,L-谷氨酰胺,通过悬浮培养,hl-60细胞能持续增殖。...
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4 月 8 日,内蒙古首例医疗事故罪案在包头市中级人民法院开庭审理,此案再一次引起网友热议。2013 年,一 3 岁男童因重症细菌性肺炎并发感染性休克,加之镇静药物的不当使用致肝脏、大脑等多器官功能损坏和衰竭死亡。村卫生室医生潘耀平被认定为承担主要责任,2014 年 9 月 19 日,检察院以医疗事故罪严重不负责任对潘耀平医生批准逮捕并提起公诉。潘耀平一审被判处有
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广州辉骏生物科技有限公司在发布的辉骏生物载体构建 病毒包装 稳转株构建等年末5折低促,还有500元京东购物卡可领供应信息,浏览与辉骏生物载体构建 病毒包装 稳转株构建等年末5折低促,还有500元京东购物卡可领相关的产品或在搜索更多与辉骏生物载体构建 病毒包装 稳转株构建等年末5折低促,还有500元京东购物卡可领相关的内容。
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简介: 【HEL细胞】,细胞库拥有专业细胞实验室及技术人员,可提供复苏细胞,冻存细胞。全国统一最低价格。专业的细胞售前,最好的细胞售后,为老师们提供放心科研细胞。具体详情请致电咨询:4000-520-616详细说明: 【HEL细胞】产品特点:细胞库拥有专业细胞实验室及技术人员,可提供复苏细胞,冻存细胞。全国统一最低价格。专业的细胞售前,最好的细胞售后,
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1.将外源基因插入慢病毒载体
2.将构建完成的载体与慢病毒包装质粒混合,共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选,如果没有,可以用无限稀释法
6.获得稳转株
1、请教各位老师,
转染质粒DNA的HepG2及HUH7细胞,如何筛选稳转
细胞株?
2、有限稀释法有没有protocol?
稳转细胞系常用的方法有质粒转染和慢病毒转染进行稳定株筛选,前者操作简单,但是成功率很低,费时,假阳性高,最后可能导致几个月的工作白干了,慢病毒稳定株筛选方法比较简单,并且慢病毒本身具有稳定整合入基因组的特性,没有假阳性,慢病毒。
稳定细胞系构建后可以开展哪些实验
建立稳定细胞株,一般是根据不同基因载体中所含有的抗性标志选用相应的药物对靶...专注整体实验·服务生命科学已开展了数千个实验外包项目
转染或PEI转染293的细胞后,一种方式加筛选压力(puromycinorhygromycin),筛选后少量细胞存活,扩大培养后,发现没有阳性,另一种方式,转染后分单克隆,扩大培养后也检测不到阳性。原本转染24-72小时转染效率至少在50%,大部分可能是瞬转,但不会一个都没有整合到基因组吧,以前我用这种方法筛选成功过几株稳转株,但是最近一段时间都失败,有遇到类似情况的吗?想和大家交流交流
应该是通过病毒载体质粒上的标记基因比如说荧光的eGFP或者带嘌呤霉素抗性基因的用嘌呤霉素筛选吧!
制备成功稳定
转染细胞株后,能否再用瞬时转染的方法转入其它质粒?
真核表达载体包括腺病毒载体和慢病毒载体,在真核细胞内用于表达目的蛋白的载体都可以叫做真核载体。 腺病毒载体携带外源基因较大,可瞬时高表达。 慢病毒载体携带外源基因较小些,但可以重组入细胞基因组,稳定高表达。
1.将外源基因插入慢病毒载体
2.将构建完成的载体与慢病毒包装质粒混合,共转染靶细胞
3.收集病毒液
4.用病毒液感染靶细胞
5.用载体上带的抗生素进行筛选,如果没有,可以用无限稀释法
6.获得稳转株
稳转细胞系常用的方法有质粒转染和慢病毒转染进行稳定株筛选,前者操作简单,但是成功率很低,费时,假阳性高,最后可能导致几个月的工作白干了,慢病毒稳定株筛选方法比较简单,并且慢病毒本身具有稳定整合入基因组的特性,没有假阳性,慢病毒包装可能相对比质粒构建难度大,如果经费充足可以直接找公司包毒的。
不能,它有Neomycin基因,可以用新霉素进行稳定株筛选,获得稳定株。
可以,除了单靶点外,该技术也可以进行多靶点基因敲除。