Topflash实验方法
WebFrom that 50ng DNA, 15ng are TOPFlash plasmid (Firefly), 10 ng Renilla, and the rest empty plasmid (PCS2 or pCDNA3). Treat the cells O/N with concentrations between 1-3uM of GSK3i or with Wnt3a.... WebB, TC1 knockdown effect on TOPFLASH analysis using HEK293 cells. The Cby-induced suppression of the reporter activity was augmented as TC1-siRNA was increased to 10 …
Topflash实验方法
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Web一般而言，topflash/fopflash是比较常用的检测β-catenin的系统，原理和所有的RLU一样，都是利用转录因子可以和特异性的靶位点结合进而启动下游的虫荧光素酶基因的表达。如果 … WebTOPflash (TCF Reporter Plasmid) Transfection grade T-cell factor reporter plasmid containing 2 sets (with the second set in the reverse orientation) of 3 copies of the TCF binding site (wild type) upstream of the Thymidine Kinase minimal promoter and Luciferase open reading frame. - Find MSDS or SDS, a COA, data sheets and more information.
WebTOPflash (TCF Reporter Plasmid) Transfection grade T-cell factor reporter plasmid containing 2 sets (with the second set in the reverse orientation) of 3 copies of the TCF … WebFrom what I can understand, TOPflash is a luciferase reporter plasmid that contains two sets of 3 copies of the wild-type TCF binding regions. If the canonical Wnt signaling is activated, beta-catenin will translocate to the nucleus to associate with TCF/LEF transcription factors to activate transcription of Wnt target genes.
WebResearchers who want to detect active Wnt-ßcatenin signaling in cells have various tools available. To measure Wnt signaling, one can use a variety of Wnt reporters. The TOP-flash assay ( Molenaar 1996) is widely used and variants of TOP-Flash (SuperTop) are available from various sources including lentivirus-based reporters from the Nusse and ... WebPlasmid M51 Super 8x FOPFlash (TOPFlash mutant) from Dr. Randall Moon's lab contains the insert mutant TCF/LEF binding sites and is published in Curr Biol. 2003 Apr 15. 13(8):680-5. This plasmid is available through Addgene.
Web用western和免疫荧光验证beta－catenin和下游靶基因的变化只是基础，只是说明你转染了这个抑癌基因的确是改变了catenin通路，但是你想要证明该抑癌基因是通过catenin通路来 …
WebThe protocol I follow is as follows: Day 1 : See 0.2e6 HEK293 Top Flash cells (stable cell lines with Renilla/Luc reporters) in 250uL DMEM-10% FBS, on a 24-well plate. Day 2 : … scanning into my macbook proWeb16. máj 2024 · Technology Overview Researchers at Johns Hopkins have developed a HEK293 cell line that is a stable reporter of canonical Wnt signaling. The cell line, which contains a luciferase reporter under control of 7 LEF/TCF binding sites, utilizes the Super TOPFlash system. scanning inventory handheldWebDay 1 : See 0.2e6 HEK293 Top Flash cells (stable cell lines with Renilla/Luc reporters) in 250uL DMEM-10% FBS, on a 24-well plate. Day 2 : Change media with 0, 50, 100, 200, 300, 400 and 500ng/ml... scanning internal and external environmentWeb22. sep 2015 · 一般而言，topflash/fopflash是比较常用的检测β-catenin的系统，原理和所有的RLU一样，都是利用转录因子可以和特异性的靶位点结合进而启动下游的虫荧光素酶基 … ruby syntax referenceWeb25. máj 2024 · Transfection grade T cell factor (Tcf) reporter plasmid containing three copies of the Tcf binding site (mutated) upstream of the Thymidine Kinase (TK) minimal … scanning into a pdfWebTOPFlash reporter luciferase activities were measured. The relative luciferase activity (fold-change) was reported. * P 0.05 versus controls ( n = 5, t test). ( O – R ) Overexpression of IGF2BP3 triggers the upregulation of KIM-1, p-H3, and p16 INK4A expression in tubular cells. HKC-8 cells were infected with either control lentivirus or ... ruby symonsWeb21. feb 2012 · 具体的TOP或FOP质粒与海肾荧光素载体的比例，需要个人通过实验优化，通常取值的范围在10/1到50/1。 数据处理： 通常以TOP/FOP的比值表示。 公式如下： 转染有TOP-Flash和海肾荧光素载体的样本中采集得到的萤火虫荧光值F (Top)/采集得到的海肾荧光值R (Top) 本回答被网友采纳 92 评论 分享 举报 shovel39 2012-02-21 · TA获得超过345个 … scanning interview questions