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Biotium/Caspase-8 IETD-R110 Fluorometric and Colorimetric Assay Kit/30011-1/25-assays
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Caspase-8isthemostupstreamcaspaseintheCD95/FasapoptoticpathwayandisactivatedbythesignalingpathwaysforCD95/FasandTNF.Caspase-8FluorometricandColorimetricAssayKitprovidesasimpleassaysystemforfastandsensitivedetectionofcaspase-8activityeitherbyfluorescenceorabsorbance.Caspase-8isthemostupstreamcaspaseintheCD95/FasapoptoticpathwayandisactivatedbythesignalingpathwaysforCD95/FasandTNF.Caspase-8FluorometricandColorimetricAssayKitprovidesasimpleassaysystemforfastandsensitivedetectionofcaspase-8activityeitherbyfluorescenceorabsorbance.Thekitincludesthefluorogenicsubstrate(Ac-IETD)2-R110,theenzymeinhibitorAc-IETD-CHOasanegativecontrol,R110asastandardandoptimizedlysisandassaybuffers.Thesubstrate(Ac-IETD)2-R110containstwoIETDtetrapeptidesandiscompletelyhydrolyzedbytheenzymeinatwo-stepprocess.CleavageofthefirstIETDresultsinthemonopeptideAc-IETD-R110intermediate,whichhasabsorptionandemissionwavelengthssimilartothoseofR110(λEx/λEm=496/520nm)buthasonlyabout10%ofthefluorescenceofthelatter.HydrolysisofthesecondIETDpeptidereleasesthedyeR110,leADIngtoasubstantialfluorescenceincrease.Althoughfluorometricdetectionoftheendproductsispreferredbecauseofthesuperiorsensitivity,detectionbyabsorbanceisalsopossIBLe.Infact,theextinctioncoefficientofR110is10timeshigherthanthatofp-nitroaniline(pNA),adyecommonlyusedinchromogenicsubstrates,makingR110-basedsubstratessignificantlymoresensitivethanpNA-basedsubstrates,evenbycolorimetricdetection.Note:Whilecaspase-8preferentiallycleavestheconsensussequenceIETDcomparedtoothersubstratesequences,othercaspasessuchascaspase-3alsocancleaveIETDefficiently.Overlappingcaspasesubstraterecognitionlimitstheusefulnessofcaspasesubstratepeptidesfordistinguishingbetweendifferentcaspaseactivitiesincelllysates.KitComponents:CelllysisbufferAssaybufferEnzymesubstrate(Ac-IETD)2-R110EnzymeinhibitorAc-IETD-CHOR110Features:Fast:Fastenzymekinetics.Sensitive:Theenzymaticreactionformsintenselyyellowcoloredandhighlygreenfluorescentrhodamine110(R110)product.Versatile:Compatiblewithbothfluorometricandcolorimetricdetectionsystems.References1)Biochemistry38,13906(1999).

EB(溴化乙锭):溴化乙锭是一种高度灵敏的荧光染色剂,用于观察琼脂糖聚丙烯酰胺凝胶中的DNA。溴化乙锭用标准302nm 紫外光透射仪激发并放射出橙红色信号,可用Polaroid 底片或带CCD 成像头的凝胶成像处理系统拍摄。

 

SYBR Green I/II是一种结合于所有dsDNA双螺旋小沟区域的具有绿色激发波长的染料。在游离状态下,SYBR Green I发出微弱的荧光,但一旦与双链DNA结合后,荧光大大增强。因此,SYBR Green I的荧光信号强度与双链DNA的数量相关,可以根据荧光信号检测出PCR体系存在的双链DNA数量。SYBR Green I 的最大吸收波长约为497nm,发射波长最大约为520nm