人凝血酶TM酶联免疫检测

AMEKOTELMEKOTEL:+86-021-55785280557585281 # FAX-mail:biolzy@人凝血酶(TM)酶联免疫检测试剂盒使用说明书□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□,□□□□□□□(TM),□□□□□□□□,□□□□□□□□□用途：□□□□□□□□□□□□□□□□□□□ (TM)测定。工作原理□□□□□□□□□□□□□□□□□□□□□□□ ELISA)测定样品中□□□(TM)水平。□□□□□□□□□□ (TM)□□□□□□□□□□□□□□血酶(TM),□□；□□□,□□□□□□□□□ TM□□□□□□□□□□ -HRP结合，□□□□□□□, □□□□□□□□, □□□□□□□, □□□□□□A、B,产生蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅与样品中人凝血酶(TM)□□□□□□□□□□□□□□□□□48口配置96口配置保存说明书1份1份封板膜2□□48)2□(96)密封袋1个1个□□□□□1口481口962-8口保存标准品1600U/L0.5mlD1瓶0.5mlD1瓶2-8口保存□□□□□□3ml口1瓶6ml口1瓶2-8口保存□□□□□ -HRP3ml口1瓶6ml口1瓶2-8口保存□□□□□□□TM抗体0.5mlD1瓶1ml口1瓶2-8口保存显色剂 A液3ml口1瓶6ml口1瓶2-8口保存显色剂 B液3ml口1瓶6ml口1瓶2-8口保存终止液3ml口1瓶6ml口1瓶2-8口保存□□□□□□20ml口20□□□ 1瓶(20mM30□□□ 1瓶2-8口保存□□□□□□□□□□□□1口37口恒温箱。2口□□□□□□□□3口□□□□□□□□□□□4口蒸馏水，5口□□□□□6口吸水口注意事项1口从2-8℃取出的试剂盒， □□□□□□□□□□□□□□□ 30分钟。酶标包□□□□□□□□□,□□□□□□□□□□□□2口□□□□□□□□□□□,□□□□□□□□□,□□□□□□□3口□□□□□□□□□□□□,□□□□□□□□□□□□□□□□ .4口□□□□□□□,□□□□□□□□□□□□□□□□□5口□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□6口底物B对光敏感，避免长时间暴露于光下。洗板方法□□□□□□□□□□□： □□□□□□□□□；□□□□□1-2□□□□□□□□□□：□□□□□□□□,□□□□□□□□□□□□□□□□□标本要求1口不能检测含 NaN3□□□,□NaN3□□□□□□□□□□□ HRP）活性。2口标本采集后尽早进行提取，提取按相关文献进行，提取后应尽快进行实验。□□□□□□□□□, □□□□□□ -20℃保存，□□□□□□□□□操作程序□□□□□□： (□□□□□□□□□□□□□,□□□□□□□□□□□□□□□□□□□ )800U/L5□□□□120pl□□□□□□□□120pl□□□□□□400U/L4号标准品120pl的5□□□□□□120pl□□□□□□200U/L3号标准品120pl的4号标准品加入120pl□□□□□□100U/L2□□□□120pl的3□□□□□□120pl□□□□□□50U/L1号标准品120pl的2□□□□□□120pl□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□ □□□□□□□□□□□□□□□□□□□□□□□□□□□□,□□□□□□□□□□□□.加口：1口空白孔，空白对照孔不加样品，生物素标记的抗 TM抗体，链霉亲和素-HRP,只加显色剂 A&B和终止液，其余各步操作相同；2）标准品孔：加入标准品50ul,链霉亲和素-HRP50U1（标准品中已事先整合好生物素抗体， □□□）；3）代测样品孔：加入 样本 40ul,然后各加入 抗TM抗体10ul、链霉亲和素-HRP50U1,盖上封板膜，轻轻震荡混匀， 37口温育 60分钟。.配液：将30□□□□□□□□□□ 30倍稀释后备用。.□□：□□□□□□□,□□□□,□□,□□□□□□□,□□ 30秒后□□,□□□□ 5次，口干。.显色：□□□□□□□□ A50ul,□□□□□□ B50pl,□□□□□□, 37℃口光显色10分钟..□□：□□□□□□ 50pl,□□□□□□□□□□□□□□ □.□□：□□□□□□, 450nm□□□□□□□□□□□□□ OD值）。测定应□□□□□□ 10分□□□□□□

9.□□□□□□□□□□□□OD□□□□□□□□□□□□□□□,□□□样品的OD□□□□□□□□□□□□□□□□□□□□□□□□□□□□9.□□□□□□□□□□□□OD□□□□□□□□□□□□□□□,□□□样品的OD□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□□准备试剂，□□□□□□检测范围：30U/L1800U/L。保存： 2-8℃。有效期： 6个月（2-81）。HumanThrombin(TM)ELISAKitInstructionThiskitisonlyforscientificresearch,andshallnotbeusedasaclinicaldiagnosisofuse.PurposeThiskitallowsforthedeterminationofTMconcentrationsinHumanserum,cellculturesupernatant,andotherbiologicalfluids.PrincipleThekitassayHumanTMlevelinthesample,addHumanTMantibodytomicrotiterplatewells,afterIncubating,addBiotinylatedantiTM-antibody,thenCombinedStreptavidin-HRP,becomecomplex,afterIncubatingandwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolor,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofTMinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.MaterialsprovidedwiththekitMaterialsprovidedwiththekit48determinations96determinationsStorageUsermanual1iClosureplatemembrane22Sealedbags11Microelisastripplate112-8℃Standard：16oou/l0.5mlx1bottle0.5mlx1bottle2-8℃Standarddiluent3mM1bottle6mlx1bottle2-8℃Streptavidin-HRP3mM1bottle6mlx1bottle2-8℃BiotinylatedantiTM-antibody0.5mlx1bottle1mlx1bottle2-8℃ChromogenSolutionA3mlx1bottle6mlx1bottle2-8℃ChromogenSolutionB3mM1bottle6mlx1bottle2-8℃StopSolution3mM1bottle6mlx1bottle2-8℃washsolution(20mlX20fold)xibottle(20mlX30fold)xibottle2-8℃Materialsrequiredbutnotsupplied1.37℃incubatorStandardmicroplatereader(450nm)PrecisionpipettesandDisposablepipettetips.deionizedwater.DisposableTesttubeAbsorbentpaperImportantnotesThekittakesoutfromtherefrigerationenvironmentshouldbebalanced15-30minutesintheroomtemperature,ELISAplatescoatedifhasnotuseupafteropened,theplateshouldbestoredinSealedbag.addSamplewithsamplerEachstep,Andproofreaditsaccuracyfrequently,avoidstheexperimentalerror.Pleaseaccordingtouseinstructionstrictly,Thetestresultdeterminationmusttakethemicrotiterplatereaderasastandard.UsenewdisposalplasticpipettetipsandClosureplatemembraneforeachstandard,inordertoavoidcrosscontamination.Donotmixreagentswiththosefromotherlots.Thesubstrateevadethelightpreservation.SpecimenrequirementsextractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,andshouldbeexperimentassoonaspossibleaftertheextraction.Ifitcant,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.CantdetectthesamplewhichcontainaN3,becauseNaN3inhibitsHRPactive.Assayprocedure.Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:800U/L5Standard120诅OriginaldensityStandard+120mStandarddiluent400U/L4Standard120g15Standard+120g1Standarddiluent200U/L3Standard120g14Standard+120glStandarddiluent100U/L2Standard120gl3Standard+120glStandarddiluent50U/L1Standard120gl2Standard+120glStandarddiluent.accordingtestingSamplenumberstodefinehowmanywellsnedd,StandardandblanksuggestedDoholes..addsample：1)blankwells:(blankcomparisonwellsdontaddsampjeBiotinylatedantiTM-antibodyandStreptavidin-HRP,othereachstepoperationissame);2)Standardwells:addStandard50andStreptavidin-HRP50;l3)testingSamplewells:addsample40,tlhenaddantiTM-antibody10,lStreptavidin-HRP50,closingplatewithClosureplatemembrane,incubatefor60minat37℃..Configurateliquid:30-fold(or20-fold)washsolutiondiluted30-fold(or20-fold)withdistilledwaterandreserve..washing：UncoverClosureplatemembrane,discardLiquid,drybyswing,addwashingbuffertoeverywell,stillfor30sthendrain,repeat5times,drybypat..color：AddChromogenSolutionA