文档简介1Humantuberculosis(TB)FORRESEARCHUSEONLYAssayrange:6pg/ml-160pg/ml96determinationsPurposeThiskitallowsforthedeterminationofTBconcentrationsinHumanserum,cellculturesupernatesandotherbiologicalfluidsPrincipleoftheassayThekitassayHumanTBlevelinthesample,usePurifiedHumanTBantibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddTBtowells,CombinedTBantibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofHumanTBinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Materialsprovidedwiththekit1washsolution20ml×1bottle7StoppSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(320pg/ml)0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11Closureplatemembrane26ChromogenSolutionB6ml×1bottle12Sealedbags1Specimenrequirements21.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.2.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.Assayprocedure1.Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:160pg/ml5Standard150lOriginaldensityStandard+150lStandarddiluent80pg/ml4Standard150l5Standard+150lStandarddiluent40pg/ml3Standard150l4Standard+150lStandarddiluent20pg/ml2Standard150l3Standard+150lStandarddiluent10pg/ml1Standard150l2Standard+150lStandarddiluent2.addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40ltotestingsamplewell,thenaddtestingsample10l(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-fold(or20-fold)washsolutiondiluted30-fold(or20-fold)withdistilledwaterandreserve.5.washing:UncoverClosureplatemembrane,discardLiquid,drybyswing,addwashingbuffertoeverywell,stillfor30sthendrain,repeat5times,drybypat.6.addenzyme:AddHRP-Conjugatereagent50ltoeachwell,exceptblankwell.7.incubate:Operationwith3.8.washing:Operationwith5.9.color:AddChromogenSolutionA50ulandChromogenSolutionBtoeachwell,evadethelightpreservationfor15minat37℃10.Stopthereaction:AddStopSolution50ltoeachwell,Stopthereaction(thebluecolorchangetoyellowcolor).11.assay:takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionand3within15min.StepsdescriptionStandard,SamplediluentAddStandard,Samplediluent,incubatefor30minat37℃.Wash5time,AddHRP-Conjugatereagent,incubatefor30minat37℃.Wash5times,AddChromogenSolutionAandB,incubatefor30minat37℃.AddStoppSolutionReadabsorbanceat450nmwithin15mincalculateCalculateTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightline4regressionequationofthestandardcurvewiththestandarddensityandtheODvalue,withthesampleODvalueintheequation,calculatethesampledensity,multipliedbythedilutionfactor,theresultisthesampleactualdensity.Importantnotes1.Thekittakesoutfromtherefrigerationenvironment首ldbebalanced15-30minutesintheroomtemperature,ELISAplatescoatedifhasnotuseupafteropened,theplate首ldbestoredinSealedbag.2.washingbufferwillCrystallizationseparation,itcanbeheatedthewaterhelpsdissolvewhendilute.Washingdoesnotaffecttheresult.3.addSamplewithsamplerEachstep,Andproofreaditsaccuracyfrequently,avoidstheexperimentalerror.addsamplewithin5min,ifthenumberofsampleismuch,recommendtouseVolley.4.ifthetestingmaterialcontentisexcessivelyhigher(ThesampleODisbiggerthanthefirststandardwell),pleasediluteSample(n-fold),Pleasediluenteandmultipliedbythedilutionfactor.(×n×5).5.Closureplatemembraneonlylimitsthedisposableuse,toavoidcross-contamination.6.Thesubstrateevadethelightpreservation.7.Pleaseaccordingtouseinstructionstrictly,Thetestresultdeterminationmusttakethemicrotiterplatereaderasastandard.8.Allsamples,washingbufferandeachkindofreject首ldaccordingtoinfectivematerialprocess.9.Donotmixreagentswiththosefromotherlots.Storageandvalidity1.Storage:2-8℃.2.validity:sixmonths上海恒远生物科技有限公司(www.shhykit.com)专业提供“人结核杆菌(TB)试剂盒”,如需了解该产品的更多信息,欢迎来电来函!上海恒远主营产品/服务:ELISA试剂盒,免疫组化试剂盒,放免试剂盒,标准品,血清,抗体,培养基,细胞,生物试剂,实验耗材等公司秉承“专注品质、信守承诺、积极沟通、创新服务”的企业文化积极参与生物领域的技术创新和技术服务,力求为我国科研事业添砖加瓦。如果你想了解更多关于“人结核杆菌(TB)试剂盒”的详细信息,欢迎联系:联系人:莫小姐联系电话:021-60515410,18221290082传真:021-64881400E-mail:shhyswkj@163.comMSN:Elisa-RD@hotmail.comhttp://www.shhybio.com/ELISA试剂盒,人ELISA试剂盒,大鼠ELISA试剂盒,小鼠ELISA试剂盒http://www.shhykit.com/进口ELISA试剂盒批发,大鼠ELISA试剂盒,进口大鼠ELISA试剂盒http://www.hyswsh.com/人ELISA试剂盒,人ELISA试剂盒价格,大鼠ELISA试剂盒,大鼠ELIS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