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儀器網(wǎng)> 資料庫(kù)>人可溶性髓系細(xì)胞觸發(fā)受體-1(sTREM-1)英文說(shuō)明書(shū).pdf

人可溶性髓系細(xì)胞觸發(fā)受體-1(sTREM-1)英文說(shuō)明書(shū).pdf

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2018-11-15 10:03 1476閱讀次數(shù)

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1HumansTREM-1FORRESEARCHUSEONLYAssayrange:8pg/ml-240pg/ml96determinationsPurposeThiskitallowsforthedeterminationofsTREM-1concentrationsinHumanserum,cellculturesupernatesandotherbiologicalfluidsPrincipleoftheassayThekitassayHumansTREM-1levelinthesample,usePurifiedHumansTREM-1antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddsTREM-1towells,CombinedsTREM-1antibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofHumansTREM-1inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Materialsprovidedwiththekit1washsolution20ml×1bottle7StopSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(480pg/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:240pg/ml5Standard150μlOriginaldensityStandard+150μlStandarddiluent120pg/ml4Standard150μl5Standard+150μlStandarddiluent60pg/ml3Standard150μl4Standard+150μlStandarddiluent30pg/ml2Standard150μl3Standard+150μlStandarddiluent15pg/ml1Standard150μl2Standard+150μlStandarddiluent2.Addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(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-Conjugatereagent50μltoeachwell,exceptblankwell.7.Incubate:Operationwith3.8.Washing:Operationwith5.9.Color:AddChromogenSolutionA50ulandChromogenSolutionB50ultoeachwell,evadethelightpreservationfor10minat37℃10.Stopthereaction:AddStopSolution50μltoeachwell,Stopthereaction(thebluecolorchangetoyellowcolor).11.Assay:takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionandwithin15min.3StepsdescriptionStandard,SamplediluentAddStandard,Samplediluent,incubatefor30minat37℃.Wash5time,AddHRP-Conjugatereagent,incubatefor30minat37℃.Wash5times,AddChromogenSolutionAandB,incubatefor10minat37℃.AddStopSolutionReadabsorbanceat450nmwithin15mincalculateCalculateTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,withthe4sampleODvalueintheequation,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上海恒遠(yuǎn)生物,專業(yè)生產(chǎn)代理各種elisa試劑盒,質(zhì)量保證,值得信賴!如果你想了解更多關(guān)于試劑盒的詳細(xì)信息,歡迎來(lái)電來(lái)函!咨詢電話:021-60515410,18221290082!

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