TaqMan protein Assay

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1、NewOneStepRT-PCRKits:TaqMan蛋白表达分析研究方案荧光定量PCR——从核酸到蛋白NorthernblotsouthernblotRealtimePCRYes？westernblot2起始样本只需10-500cells或者组织样本的1-1000ng蛋白提取物•结合抗体检测的准确性，和荧光定量PCR的灵敏度和精确度•检测细胞或组织样本里的蛋白质，定量—无需蛋白纯化步骤•应用:—微量样品蛋白检测—ＲＮＡ和蛋白表达的关联性研究>miRNA:protein>mRNA:protein—验证siRNA干扰效果—验证基因转染效果—检测FFPE&冰冻肿瘤欺骗—蛋白－蛋白相互作用3TaqM

2、an®ProteinAssaysUtilizeProximityLigationAssayTechnology1234BindingofProbestoLigationofOligosinTaqMan®RQDataAnalysisProteinTargetsProximityReal-timePCRHomogeneousassaybasedontwoantibodies,eachconjugatedtoadifferentoligonucleotide(one3’oligo,one5’oligo)WhenthetwoconjugatedantibodiesbindandareinWhent

3、hetwoconjugatedantibodiesbindandareincloseproximitytheoligonucleotidescloseproximity,theoligonucleotidescanbeligated,servingasthetemplateforreal-timePCRamplificationandquantificationInventedbyProfessorUlfLandegren,SimonFredrikssonandcolleaguesatUppsalaUniversity/OlinkBiosciencesInventedbyProfessorUl

4、fLandegren,SimonFredrikssonandcolleaguesatUppsalaUniversity/OlinkBiosciencesFredriksson,S.,etal.NatureBiotechnology20,473-477(2002)4TaqMan®蛋白表达分析试剂盒试验流程从样品制备到得到结果~3.5小时手工操作时间～1.5小时连接/TTM®FtaqMan®Fast细胞裂解抗体结合数据分析灭活Real-timePCR12345蛋白表达样本制备TaqMan蛋白表达分TaqMan蛋白表达核AppliedBiosystems数据分析试剂析试剂心试剂快速Real-Time

5、PCR仪器新鲜细通用TaqMan探胞针+快速反应试剂-总蛋白Platetransfer30min.30min.60min.60min.10min.10min.5min.5min.45min.45min.6种预先设计的TaqMan蛋白表达分析试剂盒：4种干细胞多功能性marker（人OCT3/4，人SOX2，人NANOG，人LIN28）2种对照Marker(人ICAM1，人CSTB)5TaqMan®ProteinAssaysProducts1.Pre-designedAssays(readytoaddtocell/tissuelysate)-6种预先设计的TaqMan蛋白表达分析试剂盒：-4种干

6、细胞多功能性marker（人OCT3/4，人SOX2，人NANOG，人LIN28）-2种对照Marker(人ICAM1，人CSTB)3’BBBAssayPre‐designedAssay=2Antibody‐OligoAssayProbeAProbes(linkedthroughstreptavidin‐biotin)BBBAssayProbeB2.OpenKitOpenKit–‘ToolkitToolkit’tobuildassayprobestoyourproteinofinteresttobuildassayprobestoyourproteinofinterestfromStrepta

7、vidin-Oligos(prox-oligos)&customer-suppliedbiotinylatedantibodies2streptavidinoligos(32streptavidinoligos(3’and5and5’prox-oligos)3’Prox‐OligoAProx‐OligoB6新的开放式Taqman蛋白表达试剂盒OpenKitComponents•OligoProb