Porcine IL-2
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Normal07.8磅02falsefalsefalseMicrosoftInternetExplorer4RDPorcineIL-2FORRESEARCHUSEONLYAssayrange:20pg/ml-480pg/ml96determinationsPurposeThiskitallowsforthedeterminationofIL-2concentrationsinPorcineserum,cellculturesupernatesandotherbiologicalfluidsPrincipleoftheassayThekitassayPorcineIL-2levelinthesample,usePurifiedPorcineIL-2antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddPorcineIL-2towells,CombinedantibodywhichWithHRPlabeledgoatanti-Porcinebecomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofPorcineIL-2inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Materialsprovidedwiththekit1washsolution20ml×1bottle7StoppSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(960pg/ml)0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11Closureplatemembrane26ChromogenSolutionB6ml×1bottle12Sealedbags1Specimenrequirements1.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.2.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.Assayprocedure1.Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:480pg/ml5Standard150μlOriginaldensityStandard+150μlStandarddiluent240pg/ml4Standard150μl5Standard+150μlStandarddiluent120pg/ml3Standard150μl4Standard+150μlStandarddiluent60pg/ml2Standard150μl3Standard+150μlStandarddiluent30pg/ml1Standard150μl2Standard+150μlStandarddiluent2.addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-foldwashsolutiondiluted30-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:AddChromogenSolutionA50ulandChromogenSolutionBtoeachwell,evadethelightpreservationfor15minat37℃10.Stopthereaction:AddStopSolution50μltoeachwell,Stopthereaction(thebluecolorchangetoyellowcolor).11.assay:takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionandwithin15min.StepsdescriptionStandard,SamplediluentAddStandard,Samplediluent,incubatefor30minat37℃.Wash5time,AddHRP-Conjugatereagent,incubatefor30minat37℃.Wash5times,AddChromogenSolutionAandB,incubatefor30minat37℃.AddStoppSolutionReadabsorbanceat450nmwithin15mincalculateCalculateTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,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.
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Porcine IL-2
- Normal07.8磅02falsefalsefalseMicrosoftInternetExplorer4RDPorcineIL-2FORRESEARCHUSEONLYAssayrange:20pg/ml-480pg/ml96determinationsPurposeThiskitallowsforthedeterminationofIL-2concentrationsinPorcineserum,cellculturesupernatesandotherbiologicalfluidsPrincipleoftheassayThekitassayPorcineIL-2levelinthesample,usePurifiedPorcineIL-2antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddPorcineIL-2towells,CombinedantibodywhichWithHRPlabeledgoatanti-Porcinebecomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofPorcineIL-2inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Materialsprovidedwiththekit1washsolution20ml×1bottle7StoppSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(960pg/ml)0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11Closureplatemembrane26ChromogenSolutionB6ml×1bottle12Sealedbags1Specimenrequirements1.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.2.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.Assayprocedure1.Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:480pg/ml5Standard150μlOriginaldensityStandard+150μlStandarddiluent240pg/ml4Standard150μl5Standard+150μlStandarddiluent120pg/ml3Standard150μl4Standard+150μlStandarddiluent60pg/ml2Standard150μl3Standard+150μlStandarddiluent30pg/ml1Standard150μl2Standard+150μlStandarddiluent2.addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-foldwashsolutiondiluted30-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:AddChromogenSolutionA50ulandChromogenSolutionBtoeachwell,evadethelightpreservationfor15minat37℃10.Stopthereaction:AddStopSolution50μltoeachwell,Stopthereaction(thebluecolorchangetoyellowcolor).11.assay:takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionandwithin15min.StepsdescriptionStandard,SamplediluentAddStandard,Samplediluent,incubatefor30minat37℃.Wash5time,AddHRP-Conjugatereagent,incubatefor30minat37℃.Wash5times,AddChromogenSolutionAandB,incubatefor30minat37℃.AddStoppSolutionReadabsorbanceat450nmwithin15mincalculateCalculateTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,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.[詳細(xì)]
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2024-09-29 07:02
產(chǎn)品樣冊
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Porcine IL-2試劑盒使用說明書
- Normal07.8磅02falsefalsefalseMicrosoftInternetExplorer4RDPorcineIL-2FORRESEARCHUSEONLYAssayrange:20pg/ml-480pg/ml96determinationsPurposeThiskitallowsforthedeterminationofIL-2concentrationsinPorcineserum,cellculturesupernatesandotherbiologicalfluidsPrincipleoftheassayThekitassayPorcineIL-2levelinthesample,usePurifiedPorcineIL-2antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddPorcineIL-2towells,CombinedantibodywhichWithHRPlabeledgoatanti-Porcinebecomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofPorcineIL-2inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Materialsprovidedwiththekit1washsolution20ml×1bottle7StoppSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(960pg/ml)0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11Closureplatemembrane26ChromogenSolutionB6ml×1bottle12Sealedbags1Specimenrequirements1.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.2.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.Assayprocedure1.Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:480pg/ml5Standard150μlOriginaldensityStandard+150μlStandarddiluent240pg/ml4Standard150μl5Standard+150μlStandarddiluent120pg/ml3Standard150μl4Standard+150μlStandarddiluent60pg/ml2Standard150μl3Standard+150μlStandarddiluent30pg/ml1Standard150μl2Standard+150μlStandarddiluent2.addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-foldwashsolutiondiluted30-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:AddChromogenSolutionA50ulandChromogenSolutionBtoeachwell,evadethelightpreservationfor15minat37℃10.Stopthereaction:AddStopSolution50μltoeachwell,Stopthereaction(thebluecolorchangetoyellowcolor).11.assay:takeblankwellaszero,Readabsorbanceat450nmafterAddingStopSolutionandwithin15min.StepsdescriptionStandard,SamplediluentAddStandard,Samplediluent,incubatefor30minat37℃.Wash5time,AddHRP-Conjugatereagent,incubatefor30minat37℃.Wash5times,AddChromogenSolutionAandB,incubatefor30minat37℃.AddStoppSolutionReadabsorbanceat450nmwithin15mincalculateCalculateTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,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.[詳細(xì)]
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2018-11-16 10:02
產(chǎn)品樣冊
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ELISA檢測試劑盒使用說明書,豬(Porcine)白細(xì)胞介素2(IL-2)
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本試劑盒只能用于科學(xué)研究,不得用于醫(yī)學(xué)診斷檢測原理試劑盒采用雙抗體一步夾心法酶聯(lián)免疫吸附試驗(ELISA)。往預(yù)先包被白細(xì)胞介素2(IL-2)抗體的包被微孔中,依次加入標(biāo)本、標(biāo)準(zhǔn)品、HRP標(biāo)記的檢測抗體,經(jīng)過溫育并徹底洗滌。用底物TMB顯色,TMB在過氧化物酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的白細(xì)胞介素2(IL-2)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(OD值),計算樣品濃度。樣品收集、處理及保存方法1.血清:使用不含熱原和內(nèi)毒素的試管,操作過程中避免任何細(xì)胞刺激,收集血液后,3000轉(zhuǎn)離心10分鐘將血清和紅細(xì)胞迅速小心地分離。2.血漿:EDTA、檸檬酸鹽或肝素抗凝。3000轉(zhuǎn)離心30分鐘取上清。3.細(xì)胞上清液:3000轉(zhuǎn)離心10分鐘去除顆粒和聚合物。4.組織勻漿:將組織加入適量生理鹽水搗碎。3000轉(zhuǎn)離心10分鐘取上清。5.保存:如果樣本收集后不及時檢測,請按一次用量分裝,凍存于-20℃,避免反復(fù)凍融,在室溫下解凍并確保樣品均勻地充分解凍。自備物品1.酶標(biāo)儀(450nm)2.高精度加樣器及槍頭:0.5-10uL、2-20uL、20-200uL、200-1000uL3.37℃恒溫箱操作注意事項1.試劑盒保存在2-8℃,使用前室溫平衡20分鐘。從冰箱取出的濃縮洗滌液會有結(jié)晶,這屬于正?,F(xiàn)象,水浴加熱使結(jié)晶完全溶解后再使用。2.實驗中不用的板條應(yīng)立即放回自封袋中,密封(低溫干燥)保存。3.濃度為0的S0號標(biāo)準(zhǔn)品即可視為陰性對照或者空白;按照說明書操作時樣本已經(jīng)稀釋5倍,Z終結(jié)果乘以5才是樣本實際濃度。4.嚴(yán)格按照說明書中標(biāo)明的時間、加液量及順序進(jìn)行溫育操作。5.所有液體組分使用前充分搖勻。試劑盒組成名稱96孔配置48孔配置備注微孔酶標(biāo)板12孔×8條12孔×4條無標(biāo)準(zhǔn)品0.3mL*6管0.3mL*6管無樣本稀釋液6mL3mL無檢測抗體-HRP10mL5mL無20×洗滌緩沖液25mL15mL按說明書進(jìn)行稀釋底物A6mL3mL無底物B6mL3mL無終止液6mL3mL無封板膜2張2張無說明書1份1份無自封袋1個1個無注:標(biāo)準(zhǔn)品(S0-S5)濃度依次為:0、20、40、80、160、320pg/mL試劑的準(zhǔn)備20×洗滌緩沖液的稀釋:蒸餾水按1:20稀釋,即1份的20×洗滌緩沖液加19份的蒸餾水。洗板方法1.手工洗板:甩盡孔內(nèi)液體,每孔加滿洗滌液,靜置1min后甩盡孔內(nèi)液體,在吸水紙上拍干,如此洗板5次。2.自動洗板機:每孔注入洗液350μL,浸泡1min,洗板5次。操作步驟1.從室溫平衡20min后的鋁箔袋中取出所需板條,剩余板條用自封袋密封放回4℃。2.設(shè)置標(biāo)準(zhǔn)品孔和樣本孔,標(biāo)準(zhǔn)品孔各加不同濃度的標(biāo)準(zhǔn)品50μL;3.樣本孔先加待測樣本10μL,再加樣本稀釋液40μL;空白孔不加。4.除空白孔外,標(biāo)準(zhǔn)品孔和樣本孔中每孔加入辣根過氧化物酶(HRP)標(biāo)記的檢測抗體100μL,用封板膜封住反應(yīng)孔,37℃水浴鍋或恒溫箱溫育60min。5.棄去液體,吸水紙上拍干,每孔加滿洗滌液,靜置1min,甩去洗滌液,吸水紙上拍干,如此重復(fù)洗板5次(也可用洗板機洗板)。6.每孔加入底物A、B各50μL,37℃避光孵育15min。7.每孔加入終止液50μL,15min內(nèi),在450nm波長處測定各孔的OD值。結(jié)果判斷繪制標(biāo)準(zhǔn)曲線:在Excel工作表中,以標(biāo)準(zhǔn)品濃度作橫坐標(biāo),對應(yīng)OD值作縱坐標(biāo),繪制出標(biāo)準(zhǔn)品線性回歸曲線,按曲線方程計算各樣本濃度值。試劑盒性能1.準(zhǔn)確性:標(biāo)準(zhǔn)品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值,大于等于0.9900。2.靈敏度:Zdi檢測濃度小于1.0pg/mL。3.特異性:不與其它可溶性結(jié)構(gòu)類似物交叉反應(yīng)。4.重復(fù)性:板內(nèi)、板間變異系數(shù)均小于15%。5.貯藏:2-8℃,避光防潮保存。6.有效期:6個月免責(zé)聲明1.試劑盒僅供研究使用,不得用于臨床實驗或人體實驗,否則所產(chǎn)生的一切后果,由實驗者承擔(dān),本公司概不負(fù)責(zé)。2.嚴(yán)格按照說明書操作,實驗者違反說明書操作,后果由實驗者承擔(dān)。FORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.PorcineInterleukin2(IL-2)ELISAKitinstructionIntendeduseThisIL-2ELISAkitisintendedLaboratoryforResearchuseonlyandisnotforuseindiagnosticortherapeuticprocedures.TheStopSolutionchangesthecolorfrombluetoyellowandtheintensityofthecolorismeasuredat450nmusingaspectrophotometer.InordertomeasuretheconcentrationofIL-2inthesample,thisIL-2ELISAKitincludesasetofcalibrationstandards.ThecalibrationstandardsareassayedatthesametimeasthesamplesandallowtheoperatortoproduceastandardcurveofOpticalDensityversusIL-2concentration.TheconcentrationofIL-2inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.SamplecollectionandstoragesSerum-Useaserumseparatortubeandallowsamplestoclotfor30minutesbeforecentrifugationfor10minutesatapproximately3000×g.Removeserumandassayimmediatelyoraliquotandstoresamplesat-20℃or-80℃.Avoidrepeatedfreeze-thawcyclesPlasma-CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor30minutesat3000×gat2-8℃within30minutesofcollection.Storesamplesat-20℃or-80℃.Avoidrepeatedfreeze-thawcycles.Cellculturesupernatesandotherbiologicalfluids-Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat-20℃or-80℃.Avoidrepeatedfreeze-thawcycles.Note:Thesamples首lebecentrifugateddequatelyandnohemolysisorgranulewasallowed.Materialsrequiredbutnotsupplied1.Standardmicroplatereader(450nm)2.PrecisionpipettesandDisposablepipettetips.3.37℃incubatorPrecautions1.Donotsubstitutereagentsfromonekittoanother.Standard,conjugateandmicroplatesarematchedforoptimalperformance.Useonlythereagentssuppliedbymanufacturer.2.Donotremovemicroplatefromthestoragebaguntilneeded.Unusedstrips首ldbestoredat2-8°Cintheirpouchwiththedesiccantprovided.3.Mixallreagentsbeforeusing.Removeallkitreagentsfromrefrigeratorandallowthemtoreachroomtemperature(20-25°C)MaterialssuppliedName96determinations48determinationsMicroelisastripplate12*8strips12*4stripsStandard0.3ml*6tubes0.3ml*6tubesSampleDiluent6.0ml3.0mlHRP-Conjugatereagent10.0ml5.0ml20XWashsolution25ml15mlChromogenSolutionA6.0ml3.0mlChromogenSolutionB6.0ml3.0mlStopSolution6.0ml3.0mlClosureplatemembrane22Usermanual11Sealedbags11Note:Standard(S0→S5)concentrationwasfollowedby:0,20,40,80,160,320pg/ml.Reagentpreparation20×washsolution:DilutewithDistilledordeionizedwater1:20.Assayprocedure1.Prepareallreagentsbeforestartingassayprocedure.ItisrecommendedthatallStandardsandSamplesbeaddedinduplicatetotheMicroelisaStripplate.2.Addstandard:SetStandardwells,testingsamplewells.Addstandard50μltostandardwell.3.AddSample:Addtestingsample10μlthenaddSampleDiluent40μltotestingsamplewell;Blankwelldoesn’taddanyting.4.Add100μlofHRP-conjugatereagenttoeachwell,coverwithanadhesivestripandincubatefor60minutesat37°C.5.Aspirateeachwellandwash,repeatingtheprocessfourtimesforatotaloffivewashes.Washbyfil領(lǐng)eachwellwithWashSolution(400μl)usingasquirtbottle,manifolddispenserorautowasher.Completeremovalofliquidateachstepisessentialtogoodperformance.Afterthelastwash,removeanyremainingWashSolutionbyaspiratingordecanting.Inverttheplateandblotitagainstcleanpapertowels.6.AddchromogensolutionA50μlandchromogensolutionB50μltoeachwell.Gentlymixandincubatefor15minutesat37°C.Protectfromlight.7.Add50μlStopSolutiontoeachwell.Thecolorinthewells首ldchangefrombluetoyellow.Ifthecolorinthewellsisgreenorthecolorchangedoesnotappearuniform,gentlytaptheplatetoensurethoroughmixing.8.ReadtheOpticalDensity(O.D.)at450nmusingamicrotiterplatereaderwithin15minutes.Calculationofresults1.Thisstandardcurveisusedtodeterminetheamountinanunknownsample.ThestandardcurveisgeneratedbyplottingtheaverageO.D.(450nm)obtainedforeachofthesixstandardconcentrationsonthevertical(Y)axisversusthecorrespondingconcentrationonthehorizontal(X)axis.2.First,calculatethemeanO.D.valueforeachstandardandsample.AllO.D.values,aresubtractedbythemeanvalueofthezerostandardbeforeresultinterpretation.Constructthestandardcurveusinggraphpaperorstatisticalsoftware.3.Todeterminetheamountineachsample,firstlocatetheO.D.valueontheY-axisandextendahorizontallinetothestandardcurve.Atthepointofintersection,drawaverticallinetotheX-axisandreadthecorrespondingconcentration.4.Anyvariationinoperator,pipettingandwashingtechnique,incubationtimeortemperature,andkitagecancausevariationinresult.Eachuser首ldobtaintheirownstandardcurve.5.Thesensitivitybythisassayis1.0pg/ml6.StandardcurveStorage:2-8℃.validity:sixmonths.FORRESEARCHUSEONLY;NOTFORTHERAPEUTICORDIAGNOSTICAPPLICATIONS!PLEASEREADTHROUGHENTIREPROCEDUREBEFOREBEGINNING![詳細(xì)]
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2018-11-15 10:00
產(chǎn)品樣冊
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人IL-2試劑盒,白介素2(IL-2)ELISA檢測試劑盒
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應(yīng)用文章
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牛白細(xì)胞介素2(IL-2)說明書
- www.biokanu.com牛白細(xì)胞介素2(IL-2)酶聯(lián)免疫分析試劑盒使用說明書本試劑僅供研究使用目的:本試劑盒用于測定牛血清,血漿,細(xì)胞上清及相關(guān)液體樣本中白細(xì)胞介素2(IL-2)的含量。實驗原理:本試劑盒應(yīng)用雙抗體夾心法測定標(biāo)本中牛白細(xì)胞介素2(IL-2)水平。用純化的牛白細(xì)胞介素2(IL-2)抗體包被微孔板,制成固相抗體,往包被單抗的微孔中依次加入白細(xì)胞介素2(IL-2),再與HRP標(biāo)記的白細(xì)胞介素2(IL-2)抗體結(jié)合,形成抗體-抗原-酶標(biāo)抗體復(fù)合物,經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的白細(xì)胞介素2(IL-2)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(OD值),通過標(biāo)準(zhǔn)曲線計算樣品中牛白細(xì)胞介素2(IL-2)濃度。試劑盒組成:試劑盒組成48孔配置96孔配置保存說明書1份1份封板膜2片(48)2片(96)密封袋1個1個酶標(biāo)包被板1×481×962-8℃保存標(biāo)準(zhǔn)品:360ng/L0.5ml×1瓶0.5ml×1瓶2-8℃保存標(biāo)準(zhǔn)品稀釋液1.5ml×1瓶1.5ml×1瓶2-8℃保存酶標(biāo)試劑3ml×1瓶6ml×1瓶2-8℃保存樣品稀釋液3ml×1瓶6ml×1瓶2-8℃保存顯色劑A液3ml×1瓶6ml×1瓶2-8℃保存顯色劑B液3ml×1瓶6ml×1瓶2-8℃保存終止液3ml×1瓶6ml×1瓶2-8℃保存濃縮洗滌液(20ml×20倍)×1瓶(20ml×30倍)×1瓶2-8℃保存樣本處理及要求:1.血清:室溫血液自然凝固10-20分鐘,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如出現(xiàn)沉淀,應(yīng)再次離心。2.血漿:應(yīng)根據(jù)標(biāo)本的要求選擇EDTA或檸檬酸鈉作為抗凝劑,混合10-20分鐘后,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)該再次離心。3.尿液:用無菌管收集,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)再次離心。胸腹水、腦脊液參照實行。4.細(xì)胞培養(yǎng)上清:檢測分泌性的成份時,用無菌管收集。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。檢測細(xì)胞內(nèi)的成份時,用PBS(PH7.2-7.4)稀釋細(xì)胞懸液,細(xì)胞濃度達(dá)到100萬/ml左右。通過反復(fù)凍融,以使細(xì)胞破壞并放出細(xì)胞內(nèi)成份。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。保存過程中如有沉淀形成,應(yīng)再次離心。5.組織標(biāo)本:切割標(biāo)本后,稱取重量。加入一定量的PBS,PH7.4。用液氮迅速冷凍保存?zhèn)溆谩?biāo)本融化后仍然保持2-8℃的溫度。加入一定量的PBS(PH7.4),用手工或勻漿器將標(biāo)本勻漿充分。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。分裝后一份待檢測,其余冷凍備用。6.標(biāo)本采集后盡早進(jìn)行提取,提取按相關(guān)文獻(xiàn)進(jìn)行,提取后應(yīng)盡快進(jìn)行實驗。若不能馬上進(jìn)行試驗,可將標(biāo)本放于-20℃保存,但應(yīng)避免反復(fù)凍融.7.不能檢測含NaN3的樣品,因NaN3YZ辣根過氧化物酶的(HRP)活性。操作步驟:標(biāo)準(zhǔn)品的稀釋與加樣:在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔,在**、第二孔中分別加標(biāo)準(zhǔn)品100μl,然后在**、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后從**孔、第二孔中各取100μl分別加到第三孔和第四孔,再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后在第三孔和第四孔中先各取50μl棄掉,再各取50μl分別加到第五、第六孔中,再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul,混勻;混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中,再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第七、第八孔中分別取50μl加到第九、第十孔中,再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第九第十孔中各取50μl棄掉。(稀釋后各孔加樣量都為50μl,濃度分別為240ng/L,160ng/L,80ng/L,40ng/L,20ng/L)。加樣:分別設(shè)空白孔(空白對照孔不加樣品及酶標(biāo)試劑,其余各步操作相同)、待測樣品孔。在酶標(biāo)包被板上待測樣品孔中先加樣品稀釋液40μl,然后再加待測樣品10μl(樣品Z終稀釋度為5倍)。加樣將樣品加于酶標(biāo)板孔底部,盡量不觸及孔壁,輕輕晃動混勻。溫育:用封板膜封板后置37℃溫育30分鐘。配液:將30(48T的20倍)倍濃縮洗滌液用蒸餾水30(48T的20倍)倍稀釋后備用。洗滌:小心揭掉封板膜,棄去液體,甩干,每孔加滿洗滌液,靜置30秒后棄去,如此重復(fù)5次,拍干。加酶:每孔加入酶標(biāo)試劑50μl,空白孔除外。溫育:操作同3。洗滌:操作同5。顯色:每孔先加入顯色劑A50μl,再加入顯色劑B50μl,輕輕震蕩混勻,37℃避光顯色15分鐘.終止:每孔加終止液50μl,終止反應(yīng)(此時藍(lán)色立轉(zhuǎn)黃色)。測定:以空白空調(diào)零,450nm波長依序測量各孔的吸光度(OD值)。測定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行。注意事項:試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用,酶標(biāo)包被板開封后如未用完,板條應(yīng)裝入密封袋中保存。濃洗滌液可能會有結(jié)晶析出,稀釋時可在水浴中加溫助溶,洗滌時不影響結(jié)果。各步加樣均應(yīng)使用加樣器,并經(jīng)常校對其準(zhǔn)確性,以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi),如標(biāo)本數(shù)量多,推薦使用排槍加樣。請每次測定的同時做標(biāo)準(zhǔn)曲線,**做復(fù)孔。如標(biāo)本中待測物質(zhì)含量過高(樣本OD值大于標(biāo)準(zhǔn)品孔**孔的OD值),請先用樣品稀釋液稀釋一定倍數(shù)(n倍)后再測定,計算時請Z后乘以總稀釋倍數(shù)(×n×5)。封板膜只限一次性使用,以避免交叉污染。底物請避光保存。嚴(yán)格按照說明書的操作進(jìn)行,試驗結(jié)果判定必須以酶標(biāo)儀讀數(shù)為準(zhǔn).所有樣品,洗滌液和各種廢棄物都應(yīng)按傳染物處理。本試劑不同批號組分不得混用。10.如與英文說明書有異,以英文說明書為準(zhǔn)。計算:以標(biāo)準(zhǔn)物的濃度為橫坐標(biāo),OD值為縱坐標(biāo),在坐標(biāo)紙上繪出標(biāo)準(zhǔn)曲線,根據(jù)樣品的OD值由標(biāo)準(zhǔn)曲線查出相應(yīng)的濃度;再乘以稀釋倍數(shù);或用標(biāo)準(zhǔn)物的濃度與OD值計算出標(biāo)準(zhǔn)曲線的直線回歸方程式,將樣品的OD值代入方程式,計算出樣品濃度,再乘以稀釋倍數(shù),即為樣品的實際濃度。(此圖僅供參考)試劑盒性能:1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.990以上。2.批內(nèi)與批見應(yīng)分別小于9%和11%檢測范圍:15ng/L-300ng/L保存條件及有效期:1.試劑盒保存:;2-8℃。2.有效期:6個月[詳細(xì)]
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2018-09-22 10:00
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大鼠白細(xì)胞介素2(IL-2)說明書
- www.biokanu.com大鼠白細(xì)胞介素2(IL-2)酶聯(lián)免疫分析試劑盒使用說明書本試劑僅供研究使用目的:本試劑盒用于測定大鼠血清,血漿,細(xì)胞上清及相關(guān)液體樣本中白細(xì)胞介素2(IL-2)的含量。實驗原理:本試劑盒應(yīng)用雙抗體夾心法測定標(biāo)本中大鼠白細(xì)胞介素2(IL-2)水平。用純化的大鼠白細(xì)胞介素2抗體包被微孔板,制成固相抗體,往包被單抗的微孔中依次加入白細(xì)胞介素2,再與HRP標(biāo)記的羊抗鼠抗體結(jié)合,形成抗體-抗原-酶標(biāo)抗體復(fù)合物,經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的IL-2呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(OD值),通過標(biāo)準(zhǔn)曲線計算樣品中大鼠白細(xì)胞介素2(IL-2)濃度。試劑盒組成:試劑盒組成48孔配置96孔配置保存說明書1份1份封板膜2片(48)2片(96)密封袋1個1個酶標(biāo)包被板1×481×962-8℃保存標(biāo)準(zhǔn)品:1800ng/L0.5ml×1瓶0.5ml×1瓶2-8℃保存標(biāo)準(zhǔn)品稀釋液1.5ml×1瓶1.5ml×1瓶2-8℃保存酶標(biāo)試劑3ml×1瓶6ml×1瓶2-8℃保存樣品稀釋液3ml×1瓶6ml×1瓶2-8℃保存顯色劑A液3ml×1瓶6ml×1瓶2-8℃保存顯色劑B液3ml×1瓶6ml×1瓶2-8℃保存終止液3ml×1瓶6ml×1瓶2-8℃保存濃縮洗滌液(20ml×20倍)×1瓶(20ml×30倍)×1瓶2-8℃保存樣本處理及要求:1.血清:室溫血液自然凝固10-20分鐘,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如出現(xiàn)沉淀,應(yīng)再次離心。2.血漿:應(yīng)根據(jù)標(biāo)本的要求選擇EDTA或檸檬酸鈉作為抗凝劑,混合10-20分鐘后,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)該再次離心。3.尿液:用無菌管收集,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)再次離心。胸腹水、腦脊液參照實行。4.細(xì)胞培養(yǎng)上清:檢測分泌性的成份時,用無菌管收集。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。檢測細(xì)胞內(nèi)的成份時,用PBS(PH7.2-7.4)稀釋細(xì)胞懸液,細(xì)胞濃度達(dá)到100萬/ml左右。通過反復(fù)凍融,以使細(xì)胞破壞并放出細(xì)胞內(nèi)成份。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。保存過程中如有沉淀形成,應(yīng)再次離心。5.組織標(biāo)本:切割標(biāo)本后,稱取重量。加入一定量的PBS,PH7.4。用液氮迅速冷凍保存?zhèn)溆?。?biāo)本融化后仍然保持2-8℃的溫度。加入一定量的PBS(PH7.4),用手工或勻漿器將標(biāo)本勻漿充分。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。分裝后一份待檢測,其余冷凍備用。6.標(biāo)本采集后盡早進(jìn)行提取,提取按相關(guān)文獻(xiàn)進(jìn)行,提取后應(yīng)盡快進(jìn)行實驗。若不能馬上進(jìn)行試驗,可將標(biāo)本放于-20℃保存,但應(yīng)避免反復(fù)凍融.7.不能檢測含NaN3的樣品,因NaN3YZ辣根過氧化物酶的(HRP)活性。操作步驟:1.標(biāo)準(zhǔn)品的稀釋與加樣:在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔,在**、第二孔中分別加標(biāo)準(zhǔn)品100μl,然后在**、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后從**孔、第二孔中各取100μl分別加到第三孔和第四孔,再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后在第三孔和第四孔中先各取50μl棄掉,再各取50μl分別加到第五、第六孔中,再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul,混勻;混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中,再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第七、第八孔中分別取50μl加到第九、第十孔中,再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第九第十孔中各取50μl棄掉。(稀釋后各孔加樣量都為50μl,濃度分別為1200ng/L,800ng/L,400ng/L,200ng/L,100ng/L)。2.加樣:分別設(shè)空白孔(空白對照孔不加樣品及酶標(biāo)試劑,其余各步操作相同)、待測樣品孔。在酶標(biāo)包被板上待測樣品孔中先加樣品稀釋液40μl,然后再加待測樣品10μl(樣品Z終稀釋度為5倍)。加樣將樣品加于酶標(biāo)板孔底部,盡量不觸及孔壁,輕輕晃動混勻。3.溫育:用封板膜封板后置37℃溫育30分鐘。4.配液:將30(48T的20倍)倍濃縮洗滌液用蒸餾水30(48T的20倍)倍稀釋后備用。5.洗滌:小心揭掉封板膜,棄去液體,甩干,每孔加滿洗滌液,靜置30秒后棄去,如此重復(fù)5次,拍干。6.加酶:每孔加入酶標(biāo)試劑50μl,空白孔除外。7.溫育:操作同3。8.洗滌:操作同5。9.顯色:每孔先加入顯色劑A50μl,再加入顯色劑B50μl,輕輕震蕩混勻,37℃避光顯色15分鐘.10.終止:每孔加終止液50μl,終止反應(yīng)(此時藍(lán)色立轉(zhuǎn)黃色)。11.測定:以空白空調(diào)零,450nm波長依序測量各孔的吸光度(OD值)。測定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行。注意事項:1.試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用,酶標(biāo)包被板開封后如未用完,板條應(yīng)裝入密封袋中保存。2.濃洗滌液可能會有結(jié)晶析出,稀釋時可在水浴中加溫助溶,洗滌時不影響結(jié)果。3.各步加樣均應(yīng)使用加樣器,并經(jīng)常校對其準(zhǔn)確性,以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi),如標(biāo)本數(shù)量多,推薦使用排槍加樣。4.請每次測定的同時做標(biāo)準(zhǔn)曲線,**做復(fù)孔。如標(biāo)本中待測物質(zhì)含量過高(樣本OD值大于標(biāo)準(zhǔn)品孔**孔的OD值),請先用樣品稀釋液稀釋一定倍數(shù)(n倍)后再測定,計算時請Z后乘以總稀釋倍數(shù)(×n×5)。5.封板膜只限一次性使用,以避免交叉污染。6.底物請避光保存。7.嚴(yán)格按照說明書的操作進(jìn)行,試驗結(jié)果判定必須以酶標(biāo)儀讀數(shù)為準(zhǔn).8.所有樣品,洗滌液和各種廢棄物都應(yīng)按傳染物處理。9.本試劑不同批號組分不得混用。10.如與英文說明書有異,以英文說明書為準(zhǔn)。計算:以標(biāo)準(zhǔn)物的濃度為橫坐標(biāo),OD值為縱坐標(biāo),在坐標(biāo)紙上繪出標(biāo)準(zhǔn)曲線,根據(jù)樣品的OD值由標(biāo)準(zhǔn)曲線查出相應(yīng)的濃度;再乘以稀釋倍數(shù);或用標(biāo)準(zhǔn)物的濃度與OD值計算出標(biāo)準(zhǔn)曲線的直線回歸方程式,將樣品的OD值代入方程式,計算出樣品濃度,再乘以稀釋倍數(shù),即為樣品的實際濃度。(此圖僅供參考)試劑盒性能:1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.990以上。2.批內(nèi)與批見應(yīng)分別小于9%和11%檢測范圍:80ng/L-1500ng/L保存條件及有效期:1.試劑盒保存:;2-8℃。2.有效期:6個月[詳細(xì)]
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2018-09-22 10:00
產(chǎn)品樣冊
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山羊白細(xì)胞介素2(IL-2)說明書
- 山羊白細(xì)胞介素2(IL-2)ELISA試劑盒說明書[詳細(xì)]
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2020-09-15 16:53
選購指南
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山羊白細(xì)胞介素2(IL-2)說明書
- 山羊白細(xì)胞介素2(IL-2)elisa試劑盒說明書本生(天津)健康科技有限公司供應(yīng):移液器吸嘴,ELISA試劑盒,動物血清,全系熒光定量PCR耗材,產(chǎn)品包括:PCR單管、八聯(lián)管、96孔板、384孔板。[詳細(xì)]
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2020-09-23 14:19
應(yīng)用文章
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豬白介素2(IL-2)ELISA試劑盒
- 豬白介素2(IL-2)ELISA試劑盒[詳細(xì)]
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2024-09-28 01:53
應(yīng)用文章
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人白介素2(IL-2)檢測試劑盒
- 人白介素2(IL-2)檢測試劑盒[詳細(xì)]
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2024-10-01 06:37
期刊論文
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豬白介素2(IL-2)ELISA 使用說明書
- 豬白介素2(IL-2)ELISA 使用說明書[詳細(xì)]
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2014-04-10 00:00
安裝說明
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人白細(xì)胞介素2(IL-2)檢測試劑盒
- 人白細(xì)胞介素2(IL-2)檢測試劑盒[詳細(xì)]
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2016-05-24 00:00
課件
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大鼠白介素-2(IL-2)ELISA試劑盒
- 電話:021-6533363955229872網(wǎng)址:http://www.westang.com大鼠白介素-2(IL-2)ELISA試劑盒(用于血清、血漿、細(xì)胞培養(yǎng)上清液和其它生物體液內(nèi))原理本實驗采用雙抗體夾心ABC-ELISA法。用抗大鼠IL-2單抗包被于酶標(biāo)板上,標(biāo)準(zhǔn)品和樣品中的IL-2與單抗結(jié)合,加入生物素化的抗大鼠IL-2,形成免疫復(fù)合物連接在板上,辣根過氧化物酶標(biāo)記的Streptavidin與生物素結(jié)合,加入底物工作液顯藍(lán)色,Z后加終止液硫酸,在450nm處測OD值,IL-2濃度與OD值成正比,可通過繪制標(biāo)準(zhǔn)曲線求出標(biāo)本中IL-2濃度。試劑盒組成(2-8℃保存)酶標(biāo)板(CoatedWells)96孔酶標(biāo)抗體工作液(EnzymeConjugate)12ml10×標(biāo)本稀釋液(SampleBuffer)12ml20×濃縮洗滌液(WashBuffer)50ml標(biāo)準(zhǔn)品(Standards):20ng/瓶2瓶底物工作液(TMBSolution)12ml**抗體工作液(BiotinylatedAntibody)12ml終止液(StopSolution)12ml準(zhǔn)備試劑與收集血樣1.收集標(biāo)本:血清、血漿(EDTA、檸檬酸鹽、肝素抗凝)、細(xì)胞培養(yǎng)上清液、組織勻漿等盡早檢測,2-8℃保存48小時;更長時間須冷凍(-20℃或-70℃)保存,避免反復(fù)凍融。2.標(biāo)準(zhǔn)品液配制:使用前加入1ml蒸餾水混勻,配成20ng/ml的溶液。設(shè)標(biāo)準(zhǔn)管8管,**管加標(biāo)本稀釋液900ul,第二至第八管加入標(biāo)本稀釋液500ul。在**管中加入20ng/ml的標(biāo)準(zhǔn)品溶液100ul混勻后用加樣器吸出500ul,移至第二管。如此反復(fù)作對倍稀釋,從第七管中吸出500ul棄去。第八管為空白對照。3.10×標(biāo)本稀釋液用蒸餾水作1:10倍稀釋(示例:1ml濃稀釋液+9ml蒸餾水)。4.洗滌液:用重蒸水1:20稀釋(示例:1ml濃縮洗滌液加入19ml的重蒸水)檢測程序1.加樣:每孔各加入標(biāo)準(zhǔn)品或待測樣品100ul,將反應(yīng)板充分混勻后置37℃120分鐘。2.洗板:用洗滌液將反應(yīng)板充分洗滌4-6次,向濾紙上印干。3.每孔中加入**抗體工作液100ul。將反應(yīng)板充分混勻后置37℃60分鐘。4.洗板:同前。5.每孔加酶標(biāo)抗體工作液100ul。將反應(yīng)板置37℃30分鐘。6.洗板:同前。7.每孔加入底物工作液100ul,置37℃暗處反應(yīng)15分鐘。8.每孔加入100ul終止液混勻。9.30分鐘內(nèi)用酶標(biāo)儀在450nm處測吸光值。結(jié)果計算與判斷1.所有OD值都應(yīng)減除空白值后再行計算。2.以標(biāo)準(zhǔn)品2000、1000、500、250、125、62.5、31.2、0pg/ml為橫坐標(biāo),OD值為縱坐標(biāo),在坐標(biāo)紙上作圖,畫出標(biāo)準(zhǔn)曲線。3.根據(jù)樣品OD值在該曲線圖上查出相應(yīng)IL-2含量。試劑盒性能1.靈敏度:Z小的IL-2檢測濃度小于15pg/ml。2.特異性:可同時檢測重組或天然的大鼠IL-2。不與大鼠其它細(xì)胞因子有交叉反應(yīng)。3.重復(fù)性:板內(nèi)、板見變異系數(shù)均小于9.6%。注意事項1.以上標(biāo)準(zhǔn)孔及待測樣品均建議做復(fù)孔,每次測定應(yīng)同時做標(biāo)準(zhǔn)曲線。2.洗滌過程很關(guān)鍵。洗滌不充分將導(dǎo)致極ng確度誤差及OD值錯誤地升高。3.板條開封后剩余板條要再封好,保持板條干燥。4.本試劑盒宜置4oC冰箱保存。5.本試劑盒僅用于科研,不能用于臨床診斷![詳細(xì)]
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2018-09-13 10:00
產(chǎn)品樣冊
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小鼠白細(xì)胞介素2(IL-2)試劑盒使用方法
- 檢測范圍:96T30ng/L-1200ng/L使用目的:本試劑盒用于測定小鼠血清、血漿及相關(guān)液體樣本中白細(xì)胞介素2(IL-2)含量。實驗原理本試劑盒應(yīng)用雙抗體夾心法測定標(biāo)本中小鼠白細(xì)胞介素2(IL-2)水平。用純化的小鼠白細(xì)胞介素2(IL-2)抗體包被微孔板,制成固相抗體,往包被單抗的微孔中依次加入白細(xì)胞介素2(IL-2),再與HRP標(biāo)記的白細(xì)胞介素2(IL-2)抗體結(jié)合,形成抗體-抗原-酶標(biāo)抗體復(fù)合物,經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的白細(xì)胞介素2(IL-2)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(OD值),通過標(biāo)準(zhǔn)曲線計算樣品中小鼠白細(xì)胞介素2(IL-2)濃度。試劑盒組成130倍濃縮洗滌液20ml×1瓶7終止液6ml×1瓶2酶標(biāo)試劑6ml×1瓶8標(biāo)準(zhǔn)品(2400ng/L)0.5ml×1瓶3酶標(biāo)包被板12孔×8條9標(biāo)準(zhǔn)品稀釋液1.5ml×1瓶4樣品稀釋液6ml×1瓶10說明書1份5顯色劑A液6ml×1瓶11封板膜2張6顯色劑B液6ml×1/瓶12密封袋1個[詳細(xì)]
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2018-10-08 10:01
產(chǎn)品樣冊
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小鼠(IL-2)Elisa試劑盒使用說明書
- 小鼠白細(xì)胞介素-2(IL-2)酶聯(lián)免疫分析(ELISA)試劑盒使用說明書本試劑僅供研究使用目的:本試劑盒用于測定小鼠血清,血漿及相關(guān)液體樣本中白細(xì)胞介素-2(IL-2)含量。實驗原理:本試劑盒應(yīng)用雙抗體夾心法測定標(biāo)本中小鼠白細(xì)胞介素2(IL-2)水平。用純化的小鼠白細(xì)胞介素2(IL-2)抗體包被微孔板,制成固相抗體,往包被單抗的微孔中依次加入白細(xì)胞介素2(IL-2),再與HRP標(biāo)記的白細(xì)胞介素2(IL-2)抗體結(jié)合,形成抗體-抗原-酶標(biāo)抗體復(fù)合物,經(jīng)過徹底洗滌后加底物TMB顯色。TMB在HRP酶的催化下轉(zhuǎn)化成藍(lán)色,并在酸的作用下轉(zhuǎn)化成Z終的黃色。顏色的深淺和樣品中的白細(xì)胞介素2(IL-2)呈正相關(guān)。用酶標(biāo)儀在450nm波長下測定吸光度(OD值),通過標(biāo)準(zhǔn)曲線計算樣品中小鼠白細(xì)胞介素2(IL-2)濃度。試劑盒組成:試劑盒組成48孔配置96孔配置保存說明書1份1份封板膜2片(48)2片(96)密封袋1個1個酶標(biāo)包被板1×481×962-8℃保存標(biāo)準(zhǔn)品:3600pg/mL0.5ml×1瓶0.5ml×1瓶2-8℃保存標(biāo)準(zhǔn)品稀釋液1.5ml×1瓶1.5ml×1瓶2-8℃保存酶標(biāo)試劑3ml×1瓶6ml×1瓶2-8℃保存樣品稀釋液3ml×1瓶6ml×1瓶2-8℃保存顯色劑A液3ml×1瓶6ml×1瓶2-8℃保存顯色劑B液3ml×1瓶6ml×1瓶2-8℃保存終止液3ml×1瓶6ml×1瓶2-8℃保存濃縮洗滌液(20ml×20倍)×1瓶(20ml×30倍)×1瓶2-8℃保存樣本處理及要求:1.血清:室溫血液自然凝固10-20分鐘,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如出現(xiàn)沉淀,應(yīng)再次離心。2.血漿:應(yīng)根據(jù)標(biāo)本的要求選擇EDTA、者檸檬酸鈉或肝素作為抗凝劑,混合10-20分鐘后,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)該再次離心。3.尿液:用無菌管收集,離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清,保存過程中如有沉淀形成,應(yīng)再次離心。胸腹水、腦脊液參照實行。4.細(xì)胞培養(yǎng)上清:檢測分泌性的成份時,用無菌管收集。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。檢測細(xì)胞內(nèi)的成份時,用PBS(PH7.2-7.4)稀釋細(xì)胞懸液,細(xì)胞濃度達(dá)到100萬/ml左右。通過反復(fù)凍融,以使細(xì)胞破壞并放出細(xì)胞內(nèi)成份。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。保存過程中如有沉淀形成,應(yīng)再次離心。5.組織標(biāo)本:切割標(biāo)本后,稱取重量。加入一定量的PBS,PH7.4。用液氮迅速冷凍保存?zhèn)溆谩?biāo)本融化后仍然保持2-8℃的溫度。加入一定量的PBS(PH7.4),用手工或勻漿器將標(biāo)本勻漿充分。離心20分鐘左右(2000-3000轉(zhuǎn)/分)。仔細(xì)收集上清。分裝后一份待檢測,其余冷凍備用。操作步驟1.標(biāo)準(zhǔn)品的稀釋與加樣:在酶標(biāo)包被板上設(shè)標(biāo)準(zhǔn)品孔10孔,在**、第二孔中分別加標(biāo)準(zhǔn)品100μl,然后在**、第二孔中加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后從**孔、第二孔中各取100μl分別加到第三孔和第四孔,再在第三、第四孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻;然后在第三孔和第四孔中先各取50μl棄掉,再各取50μl分別加到第五、第六孔中,再在第五、第六孔中分別加標(biāo)準(zhǔn)品稀釋液50ul,混勻;混勻后從第五、第六孔中各取50μl分別加到第七、第八孔中,再在第七、第八孔中分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第七、第八孔中分別取50μl加到第九、第十孔中,再在第九第十孔分別加標(biāo)準(zhǔn)品稀釋液50μl,混勻后從第九第十孔中各取50μl棄掉。(稀釋后各孔加樣量都為50μl,濃度分別為2400pg/mL,1600pg/mL,800pg/mL,400pg/mL,200pg/mL)。2.加樣:分別設(shè)空白孔(空白對照孔不加樣品及酶標(biāo)試劑,其余各步操作相同)、待測樣品孔。在酶標(biāo)包被板上待測樣品孔中先加樣品稀釋液40μl,然后再加待測樣品10μl(樣品Z終稀釋度為5倍)。加樣將樣品加于酶標(biāo)板孔底部,盡量不觸及孔壁,輕輕晃動混勻。3.溫育:用封板膜封板后置37℃溫育30分鐘。4.配液:將30(48T的20倍)倍濃縮洗滌液用蒸餾水30(48T的20倍)倍稀釋后備用。5.洗滌:小心揭掉封板膜,棄去液體,甩干,每孔加滿洗滌液,靜置30秒后棄去,如此重復(fù)5次,拍干。6.加酶:每孔加入酶標(biāo)試劑50μl,空白孔除外。7.溫育:操作同3。8.洗滌:操作同5。9.顯色:每孔先加入顯色劑A50μl,再加入顯色劑B50μl,輕輕震蕩混勻,37℃避光顯色15分鐘.10.終止:每孔加終止液50μl,終止反應(yīng)(此時藍(lán)色立轉(zhuǎn)黃色)。11.測定:以空白空調(diào)零,450nm波長依序測量各孔的吸光度(OD值)。測定應(yīng)在加終止液后15分鐘以內(nèi)進(jìn)行。注意事項:1.試劑盒從冷藏環(huán)境中取出應(yīng)在室溫平衡15-30分鐘后方可使用,酶標(biāo)包被板開封后如未用完,板條應(yīng)裝入密封袋中保存。2.濃洗滌液可能會有結(jié)晶析出,稀釋時可在水浴中加溫助溶,洗滌時不影響結(jié)果。3.各步加樣均應(yīng)使用加樣器,并經(jīng)常校對其準(zhǔn)確性,以避免試驗誤差。一次加樣時間**控制在5分鐘內(nèi),如標(biāo)本數(shù)量多,推薦使用排槍加樣。4.請每次測定的同時做標(biāo)準(zhǔn)曲線,**做復(fù)孔。如標(biāo)本中待測物質(zhì)含量過高(樣本OD值大于標(biāo)準(zhǔn)品孔**孔的OD值),請先用樣品稀釋液稀釋一定倍數(shù)(n倍)后再測定,計算時請Z后乘以總稀釋倍數(shù)(×n×5)。5.封板膜只限一次性使用,以避免交叉污染。6.底物請避光保存。7.嚴(yán)格按照說明書的操作進(jìn)行,試驗結(jié)果判定必須以酶標(biāo)儀讀數(shù)為準(zhǔn).8.所有樣品,洗滌液和各種廢棄物都應(yīng)按傳染物處理。9.本試劑不同批號組分不得混用。10.如與英文說明書有異,以英文說明書為準(zhǔn)。計算:以標(biāo)準(zhǔn)物的濃度為橫坐標(biāo),OD值為縱坐標(biāo),在坐標(biāo)紙上繪出標(biāo)準(zhǔn)曲線,根據(jù)樣品的OD值由標(biāo)準(zhǔn)曲線查出相應(yīng)的濃度;再乘以稀釋倍數(shù);或用標(biāo)準(zhǔn)物的濃度與OD值計算出標(biāo)準(zhǔn)曲線的直線回歸方程式,將樣品的OD值代入方程式,計算出樣品濃度,再乘以稀釋倍數(shù),即為樣品的實際濃度。(此圖僅供參考)試劑盒性能:1.樣品線性回歸與預(yù)期濃度相關(guān)系數(shù)R值為0.95以上。2.批內(nèi)與批見應(yīng)分別小于9%和11%檢測范圍:150pg/mL-3000pg/mL保存條件及有效期:1.試劑盒保存:;2-8℃。2.有效期:6個月[詳細(xì)]
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2018-10-23 10:31
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