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斑马鱼卵黄蛋白原ELISA KIT
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2024-09-28 00:19 878阅读次数
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斑马鱼卵黄蛋白原ELISA KIT
- 斑马鱼卵黄蛋白原ELISA KIT[详细]
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2024-09-28 00:19
应用文章
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鱼卵黄蛋白原(VTG)ELISA试剂盒说明书
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FishVTGELISAKitForthequantitativeinvitrodeterminationofFishVitellogeninconcentrationsinbodyfluid-celiacfluid-tissuehomogenates-otherbiologicalfluidsFORLABORATORYRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.Thispackageinsertmustbereadinitsentiretybeforeusingthisproduct.ELISAENZYMELINKEDIMMUNOSORBENTASSAYINTENDEDUSEANDTESTPRINCIPLEThisVTGELISAkitisintendedLaboratoryforResearchuseonlyandisnotforuseindiagnosticortherapeuticprocedures.TheStopSolutionchangesthecolorfrombluetoyellowandtheintensityofthecolorismeasuredat450nmusingaspectrophotometer.InordertomeasuretheconcentrationofVTGinthesample,thisVTGELISAKitincludesasetofcalibrationstandards.ThecalibrationstandardsareassayedatthesametimeasthesamplesandallowtheoperatortoproduceastandardcurveofOpticalDensityversusVTGconcentration.TheconcentrationofVTGinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.SAMPLECOLLECTIONANDSTORAGESSerum-Useaserumseparatortubeandallowsamplestoclotfortwohoursatroomtemperatureorovernightat4℃beforecentrifugationfor20minutesatapproximately1000×g.Assayfreshlypreparedserumimmediatelyorstoresamplesinaliquotat-20℃or-80℃forlateruse.Avoidrepeatedfreeze/thawcycles.Plasma-CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor15minutesat1000×gat2-8℃within30minutesofcollection.Removeplasmaandassayimmediatelyorstoresamplesinaliquotat-20℃or-80℃forlateruse.Avoidrepeatedfreeze/thawcycles.Tissuehomogenates-Forgeneralinformation,hemolysisbloodmayaffecttheresult,soyou首ldrinsethetissueswithice-coldPBS(0.01M,pH=7.4)toremoveexcessbloodthoroughly.Tissuepieces首ldbeweighedandthenmincedtosmallpieceswhichwillbehomogenizedinPBS(thevolumedependsontheweightofthetissue.9mLPBSwouldbeappropriateto1gramtissuepieces.SomeproteaseinhibitorisrecommendedtoaddintothePBS.)withaglasshomogenizeronice.Tofurtherbreakthecells,youcansonicatethesuspensionwithanultrasoniccelldisrupterorsubjectittofreeze-thawcycles.Thehomogenatesarethencentrifugatedfor5minutesat5000×gtogetthesupernate.Cellculturesupernatesandotherbiologicalfluids-Centrifugesamplesfor20minutesat1000×g.Removeparticulatesandassayimmediatelyorstoresamplesinaliquotat-20℃or-80℃forlateruse.Avoidrepeatedfreeze/thawcycles.Note:Thesamples首lebecentrifugateddequatelyandnohemolysisorgranulewasallowed.MATERIALSREQUIREDBUTNOTSUPPLIED1.37℃incubator2.Standardmicroplatereadercapableofmeasuringabsorbanceat450nm3.Precisionpipettes,disposablepipettetipsandAbsorbentpaper4.DistilledordeionizedwaterREAGENTSPROVIDEDAllreagentsprovidedarestoredat2-8°C.Refertotheexpirationdateonthelabel.Name96determinations48determinationsMICROTITERPLATE8*12strips8*6stripsSTANDARD(6vial)0.3ml/vial0.3ml/vialSAMPLEDILUENT6.0ml3.0mlENZYMECONJUGATE10.0ml5.0mlWASHSOLUTION25ml15mlSUBSTRATEA6.0ml3.0mlSUBSTRATEB6.0ml3.0mlSTOPSOLUTION6.0ml3.0mlClosureplatemembrane22Usermanual11Sealedbags11Note:1.Standardconcentrationwasfollowedby:480,240,120,60,30,15ng/mL.2.Ifsamplesgeneratevalueshigherthanthehigheststandard,pleasedilutethesampleswithSampleDiluentandrepeattheassay.PRECAUTIONSDonotsubstitutereagentsfromonekitlottoanother.Standard,conjugateandmicrotiterplatesarematchedforoptimalperformance.Useonlythereagentssuppliedbymanufacturer.Allowkitreagentsandmaterialstoreachroomtemperature(20-25°C)beforeuse.Donotusewaterbathstothawsamplesorreagents.Donotusekitcomponentsbeyondtheirexpirationdate.Useonlydeionizedordistilledwatertodilutereagents.Donotremovemicrotiterplatefromthestoragebaguntilneeded.Unusedstrips首ldbestoredat2-8°Cintheirpouchwiththedesiccantprovided.Usefreshdisposablepipettetipsforeachtransfertoavoidcontamination.Donotmixacidandsodiumhypochloritesolutions.Serumandplasma首ldbehandledaspotentiallyhazardousandcapableoftransmittingdisease.Disposableglovesmustbewornduringtheassayprocedure,sincenoknowntestmethodcanoffercompleteassurancethatproductsderivedfromRatbloodwillnottransmitinfectiousagents.Therefore,allbloodderivatives首ldbeconsideredpotentiallyinfectiousandgoodlaboratorypractices首ldbefollowed.Allsamples首ldbedisposedofinamannerthatwillinactivateviruses.LiquidWaste:Addsodiumhypochloritetoafinalconcentrationof1.0%.Thewaste首ldbeallowedtostandforaminimumof30minutestoinactivatethevirusesbeforedisposal.SubstrateSolutioniseasilycontaminated.Ifbluishpriortouse,donotuse.SubstrateBcontain20%acetone,keepthisreagentawayfromsourcesofheatorflame.Removeallkitreagentsfromrefrigeratorandallowthemtoreachroomtemperature(20-25°C).REAGENTPREPARATIONANDSTORAGEWashSolution(1X)-Dilute1volumeofWashsolution(20X)with19volumesofdeionizedordistilledwater.WashSolutionisstablefor1monthat2-8°C.ASSAYPROCEDURE1.Prepareallreagentsbeforestartingassayprocedure.ItisrecommendedthatallStandardsandSamplesbeaddedinduplicatetotheMicrotiterplate.2.Add50μlofStandardorSampletotheappropriatewells.Blankwelldoesn’taddanyting.3.Add100μlofEnzymeconjugatetostandardwellsandsamplewellsexcepttheblankwell,coverwithanadhesivestripandincubatefor60minutesat37°C.4.WashtheMicrotiterPlate4times.ManualWashing-Removeincubationmixturebyaspiratingcontentsoftheplateintoasinkorproperwastecontainer.Usingasquirtbottle,filleachwellcompletelywithWashSolution(1X),thenaspiratecontentsoftheplateintoasinkorproperwastecontainer.Repeatthisprocedureforatotaloffourtimes.Afterfinalwash,invertplate,andblotdrybyhittingplateontoabsorbentpaperorpapertowelsuntilnomoistureappears.Note:Holdthesidesoftheplateframefirmlywhenwashingtheplatetoassurethatallstripsremainsecurelyinframe.AutomatedWashing-Aspirateallwells,thenwashplatesfourtimesusingWashBuffer(1X).Alwaysadjustyourwashertoaspirateasmuchliquidaspossibleandsetfillvolumeat350μL/well/wash.Afterfinalwash,invertplate,andblotdrybyhittingplateontoabsorbentpaperorpapertowelsuntilnomoistureappears.5.AddSubstrateA50μlandSubstrateB50μltoeachwell.Gentlymixandincubatefor15minutesat37°C.Protectfromlight.6.Add50μlStopSolutiontoeachwell.Thecolorinthewells首ldchangefrombluetoyellow.Ifthecolorinthewellsisgreenorthecolorchangedoesnotappearuniform,gentlytaptheplatetoensurethoroughmixing.7.ReadtheOpticalDensity(O.D.)at450nmusingamicrotiterplatereaderwithin15minutes.CALCULATIONOFRESULTSThisstandardcurveisusedtodeterminetheamountinanunknownsample.ThestandardcurveisgeneratedbyplottingtheaverageO.D.(450nm)obtainedforeachofthesixstandardconcentrationsonthevertical(X)axisversusthecorrespondingconcentrationonthehorizontal(Y)axis.First,calculatethemeanO.D.valueforeachstandardandsample.AllO.D.Valuesaresubtractedbythemeanvalueofthebalnkwellbeforeresultinterpretation.Constructthestandardcurveusinggraphpaperorstatisticalsoftware.Todeterminetheamountineachsample,firstlocatetheO.D.valueontheY-axisandextendahorizontallinetothestandardcurve.Atthepointofintersection,drawaverticallinetotheX-axisandreadthecorrespondingconcentration.Anyvariationinoperator,pipettingandwashingtechnique,incubationtimeortemperature,andkitagecancausevariationinresult.Eachuser首ldobtaintheirownstandardcurve.Intra-assayCV(%)islessthan10%andInter-assayCV(%)islessthan15%.Assayrange:15ng/mL480ng/mL.7.Sensitivity:TheminimumdetectabledoseofFishVTGistypicallylessthan1.0ng/mL.8.Cross-reactivity:ThisassayrecognizesrecombinantandnaturalFishVTG.Nosignificantcross-reactivityorinterferencewasobserved.9.Storage:2-8℃(Usefrequently);sixmonths(-20℃)。10.StandardcurveFORRESEARCHUSEONLY;NOTFORTHERAPEUTICORDIAGNOSTICAPPLICATIONS!PLEASEREADTHROUGHENTIREPROCEDUREBEFOREBEGINNING!鱼卵黄蛋白原(VTG)试剂盒(ELISA)使用说明书l本试剂盒用于体外定量检测体液、腔液、组织匀浆及相关液体样本中鱼卵黄蛋白原(VTG)的含量。l有效期:6个月l保存条件:2-8℃实验原理试剂盒采用双抗体一步夹心法酶联免疫吸附试验(ELISA)。往预先包被鱼卵黄蛋白原(VTG)捕获抗体的包被微孔中,依次加入标本、标准品、HRP标记的检测抗体,经过温育并彻底洗涤。用底物TMB显色,TMB在过氧化物酶的催化下转化成蓝色,并在酸的作用下转化成Z终的黄色。颜色的深浅和样品中的鱼卵黄蛋白原(VTG)呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),计算样品浓度。样本处理及要求1.血清:将收集于血清分离管的全血标本在室温放置2小时或4℃过夜,然后1000×g离心20分钟,取上清即可,或将上清置于-20℃或-80℃保存,但应避免反复冻融。2.血浆:用EDTA或肝素作为抗凝剂采集标本,并将标本在采集后的30分钟内于2-8℃1000×g离心15分钟,取上清即可检测,或将上清置于-20℃或-80℃保存,但应避免反复冻融。3.组织匀浆:用预冷的PBS(0.01M,pH=7.4)冲洗组织,去除残留血液(匀浆中裂解的红细胞会影响测量结果),称重后将组织剪碎。将剪碎的组织与对应体积的PBS(一般按1:9的重量体积比,比如1g的组织样品对应9mL的PBS,具体体积可根据实验需要适当调整,并做好记录。推荐在PBS中加入蛋白酶YZ剂)加入玻璃匀浆器中,于冰上充分研磨。为了进一步裂解组织细胞,可以对匀浆液进行超声破碎,或反复冻融。Z后将匀浆液于5000×g离心5~10分钟,取上清检测。4.细胞培养物上清或其它生物标本:请1000×g离心20分钟,取上清即可检测,或将上清置于-20℃或-80℃保存,但应避免反复冻融。注:标本溶血会影响Z后检测结果,因此溶血标本不宜进行此项检测。需要而未提供的试剂和器材酶标仪(450nm)高精度加样器及枪头:0.5-10uL、2-20uL、20-200uL、200-1000uL37℃恒温箱蒸馏水或去离子水试剂盒组成名称96孔配置48孔配置备注微孔酶标板8孔×12条8孔×6条无标准品0.3mL*6管0.3mL*6管无样本稀释液6mL3mL无检测抗体-HRP10mL5mL无20×洗涤缓冲液25mL15mL按说明书进行稀释底物A6mL3mL无底物B6mL3mL无终止液6mL3mL无封板膜2张2张无说明书1份1份无自封袋1个1个无备注:1.标准品浓度依次为:480、240、120、60、30、15ng/mL2.经过大量正常标本检验,标本的正常浓度值均在试剂盒提供的检测范围内,实验过程中直接取50μL样本上样即可。当有部分样本值超过Zda标准品浓度时,可用样本稀释液将标本进行适当稀释后再进行实验。注意事项严格按照规定的时间和温度进行温育以保证准确结果。所有试剂都必须在使用前达到室温20-25℃。使用后立即冷藏保存试剂。洗板不正确可以导致不准确的结果。在加入底物前确保尽量吸干孔内液体。温育过程中不要让微孔干燥掉。消除板底残留的液体和手指印,否则影响OD值。底物显色液应呈无色或很浅的颜色,已经变蓝的底物液不能使用。避免试剂和标本的交叉污染以免造成错误结果。在储存和温育时避免强光直接照射。平衡至室温后再打开密封袋以防水滴凝聚在冷板条上。任何反应试剂不能接触漂白溶剂或漂白溶剂所散发的强烈气体。任何漂白成分都会破坏试剂盒中反应试剂的生物活性。不能使用过期产品。如果可能传播疾病,所有的样品都应管理好,按照规定的程序处理样品和检测装置。试剂准备试剂盒从冷藏环境中取出应在室温平衡后方可使用。20×洗涤缓冲液的稀释:蒸馏水按1:20稀释,即1份20×洗涤缓冲液加19份蒸馏水。操作步骤从室温平衡20min后的铝箔袋中取出所需板条,剩余板条用自封袋密封放回4℃。设置标准品孔和样本孔,标准品孔各加不同浓度的标准品50μL;样本孔中加入待测样本50μL;空白孔不加。除空白孔外,标准品孔和样本孔中每孔加入辣根过氧化物酶(HRP)标记的检测抗体100μL,用封板膜封住反应孔,37℃水浴锅或恒温箱温育60min。弃去液体,吸水纸上拍干,每孔加满洗涤液(350μL),静置1min,甩去洗涤液,吸水纸上拍干,如此重复洗板5次(也可用洗板机洗板)。每孔加入底物A、B各50μL,37℃避光孵育15min。每孔加入终止液50μL,15min内,在450nm波长处测定各孔的OD值。实验结果计算以所测标准品的OD值为横坐标,标准品的浓度值为纵坐标,在坐标纸上或用相关软件绘制标准曲线,并得到直线回归方程,将样品的OD值代入方程,计算出样品的浓度。试剂盒性能检测范围:15ng/mL480ng/mL。灵敏度:Zdi检测浓度小于1.0ng/mL。特异性:不与其它可溶性结构类似物交叉反应。重复性:板内变异系数小于10%,板间变异系数小于15%。[详细]
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2018-12-11 10:00
产品样册
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鱼卵黄蛋白原(VTG)酶联免疫分析试剂盒使用说明书
- 鱼卵黄蛋白原(VTG)酶联免疫分析试剂盒使用说明书本试剂盒仅供研究使用。检测范围:96T30μg/L-850μg/L使用目的:本试剂盒用于测定鱼血清、血浆及相关液体样本中卵黄蛋白原(VTG)含量。实验原理本试剂盒应用双抗体夹心法测定标本中鱼卵黄蛋白原(VTG)水平。用纯化的鱼卵黄蛋白原(VTG)抗体包被微孔板,制成固相抗体,往包被单抗的微孔中依次加入卵黄蛋白原(VTG),再与HRP标记的卵黄蛋白原(VTG)抗体结合,形成抗体-抗原-酶标抗体复合物,经过彻底洗涤后加底物TMB显色。TMB在HRP酶的催化下转化成蓝色,并在酸的作用下转化成Z终的黄色。颜色的深浅和样品中的卵黄蛋白原(VTG)呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),通过标准曲线计算样品中鱼卵黄蛋白原(VTG)浓度。鱼卵黄蛋白原(VTG)酶联免疫分析试剂盒组成标本要求1.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。若不能马上进行试验,可将标本放于-20℃保存,但应避免反复冻融2.不能检测含NaN3的样品,因NaN3YZ辣根过氧化物酶的(HRP)活性。操作步骤1.标准品的稀释:本试剂盒提供原倍标准品一支,用户可按照下列图表在小试管中进行稀释。2.加样:分别设空白孔(空白对照孔不加样品及酶标试剂,其余各步操作相同)、标准孔、待测样品孔。在酶标包被板上标准品准确加样50μl,待测样品孔中先加样品稀释液40μl,然后再加待测样品10μl(样品Z终稀释度为5倍)。加样将样品加于酶标板孔底部,尽量不触及孔壁,轻轻晃动混匀。3.温育:用封板膜封板后置37℃温育30分钟。4.配液:将30倍浓缩洗涤液用蒸馏水30倍稀释后备用5.洗涤:小心揭掉封板膜,弃去液体,甩干,每孔加满洗涤液,静置30秒后弃去,如此重复5次,拍干。6.加酶:每孔加入酶标试剂50μl,空白孔除外。7.温育:操作同3。8.洗涤:操作同5。9.显色:每孔先加入显色剂A50μl,再加入显色剂B50μl,轻轻震荡混匀,37℃避光显色15分钟.10.终止:每孔加终止液50μl,终止反应(此时蓝色立转黄色)。11.测定:以空白空调零,450nm波长依序测量各孔的吸光度(OD值)。测定应在加终止液后15分钟以内进行。鱼卵黄蛋白原(VTG)酶联免疫分析试剂盒操作程序总结:计算以标准物的浓度为横坐标,OD值为纵坐标,在坐标纸上绘出标准曲线,根据样品的OD值由标准曲线查出相应的浓度;再乘以稀释倍数;或用标准物的浓度与OD值计算出标准曲线的直线回归方程式,将样品的OD值代入方程式,计算出样品浓度,再乘以稀释倍数,即为样品的实际浓度。注意事项1.试剂盒从冷藏环境中取出应在室温平衡15-30分钟后方可使用,酶标包被板开封后如未用完,板条应装入密封袋中保存。2.浓洗涤液可能会有结晶析出,稀释时可在水浴中加温助溶,洗涤时不影响结果。3.各步加样均应使用加样器,并经常校对其准确性,以避免试验误差。一次加样时间**控制在5分钟内,如标本数量多,推荐使用排枪加样。4.请每次测定的同时做标准曲线,**做复孔。如标本中待测物质含量过高(样本OD值大于标准品孔**孔的OD值),请先用样品稀释液稀释一定倍数(n倍)后再测定,计算时请Z后乘以总稀释倍数(×n×5)。5.封板膜只限一次性使用,以避免交叉污染。6.底物请避光保存。7.严格按照说明书的操作进行,试验结果判定必须以酶标仪读数为准.8.所有样品,洗涤液和各种废弃物都应按传染物处理。9.本试剂不同批号组分不得混用。鱼卵黄蛋白原(VTG)酶联免疫分析试剂盒保存条件及有效期1.试剂盒保存:2-8℃。2.有效期:6个月[详细]
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2018-11-04 10:00
产品样册
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鲤鱼卵黄蛋白原ELISA检测试剂盒
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2024-10-05 20:57
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猪血纤蛋白原(Fbg)ELISA试剂盒
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Monkey ELISA Kit
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兔血纤蛋白原(Fbg)ELISA试剂盒
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人凝血酶ELISA Kit
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Human PLA2R1 ELISA kit
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Rat Aβ1-42 ELISA kit
- www.biokanu.comRatamyloidbetapeptide1-42(Aβ1-42)ELISAKitProd.No.3R285FROM:RBForthequantitativeinvitrodeterminationofAβ1-42concentrationsinRatsupernates,serum,plasmaandtissue.FORRESEARCHUSEONLY.NOTFORUSEINDIAGNOSTICPROCEDURES.TABLEOFCONTENTSContentsPageTABLEOFCONTENTS..2INTENDEDUSE..3PRINCIPLE..3WARNINGSANDPRECAUTIONS..4MATERIALSPROVIDEDWITHTHEKIT.7MATERIALSREQUIREDBUTNOTPROVIDED..7STORAGECONDITIONS..8REAGENTPREPARATION..9SPECIMENCOLLECTIONANDPREPARATION..9ASSAYPROCEDURE..10CALCULATIONOFRESULTS..13REFERENCES..14INTENDEDUSEAnenzymeimmunoassayforthequantitativeinvitrodiagnosticmeasurementofRatAβ1-42incellculturesupernates,serum,plasmaandtissue.PRINCIPLEThekitassayRatAβ1-42levelinthesample,usePurifiedRatAβ1-42antibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddAβ1-42towells,CombinedAβ1-42antibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofAβ1-42inthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.WARNINGSANDPRECAUTIONSlThiskitisforinvitrodiagnosticuseonly.Forprofessionaluseonly.lAllreagentsofthistestkitwhichcontainhumanserumorplasmahavebeentestedandconfirmednegativeforHIVI/II,HBsAgandHCVbyFDAapprovedprocedures.Allreagents,however,首ldbetreatedaspotentialbiohazardsinuseandfordisposal.lBeforestartingtheassay,readtheinstructionscompletelyandcarefully.Usethevalidversionofthepackageinsertprovidedwiththekit.Besurethateverythingisunderstood.lThemicroplatecontainssnap-offstrips.Unusedwellsmustbestoredat2°Cto8°Cinthesealedfoilpouchandusedintheframeprovided.lPipettingofsamplesandreagentsmustbedoneasquicklyaspossibleandinthesamesequenceforeachstep.lUsereservoirsonlyforsinglereagents.Thisespeciallyappliestothesubstratereservoirs.Usingareservoirfordispensingasubstratesolutionthathadpreviouslybeenusedfortheconjugatesolutionmayturnsolutioncolored.Donotpourreagentsbackintovialsasreagentcontaminationmayoccur.lMixthecontentsofthemicroplatewellsthoroughlytoensuregoodtestresults.Donotreusemicrowells.lDonotletwellsdryduringassay;addreagentsimmediatelyaftercompletingtherinsingsteps.lAllowthereagentstoreachroomtemperature(21-26°C)beforestartingthetest.Temperaturewillaffecttheabsorbancereadingsoftheassay.However,valuesforthepatientsampleswillnotbeaffected.lNeverpipetbymouthandavoidcontactofreagentsandspecimenswithskinandmucousmembranes.lDonotsmoke,eat,drinkorapplycosmeticsinareaswherespecimensorkitreagentsarehandled.lWeardisposablelatexgloveswhenhand领specimensandreagents.Microbialcontaminationofreagentsorspecimensmaygivefalseresults.lHand领首ldbedoneinaccordancewiththeproceduresdefinedbyanappropriatenationalbiohazardsafetyguidelineorregulation.lDonotusereagentsbeyondexpirydateasshownonthekitlabels.lAllindicatedvolumeshavetobeperformedaccordingtotheprotocol.Optimaltestresultsareonlyobtainedwhenusingcalibratedpipettesandmicrotiterplatereaders.lDonotmixorusecomponentsfromkitswithdifferentlotnumbers.Itisadvisednottoexchangewellsofdifferentplatesevenofthesamelot.Thekitsmayhavebeenshippedorstoredunderdifferentconditionsandthebindingcharacteristicsoftheplatesmayresultslightlydifferent.lAvoidcontactwithStopSolutioncontaining0.5MH2SO4.Itmaycauseskinirritationandburns.lSomereagentscontainProclin,BNDand/orMITaspreservatives.Incaseofcontactwitheyesorskin,flushimmediatelywithwater.lTMBsubstratehasanirritanteffectonskinandmucosa.Incaseofpossiblecontact,washeyeswithanabundantvolumeofwaterandskinwithsoapandabundantwater.Washcontaminatedobjectsbeforereusingthem.Ifinhaled,takethepersontoopenair.lChemicalsandpreparedorusedreagentshavetobetreatedashazardouswasteaccordingtothenationalbiohazardsafetyguidelineorregulation.lForinformationonhazardoussubstancesincludedinthekitpleaserefertoMaterialSafetyDataSheetsMATERIALSPROVIDEDWITHTHEKITMaterialsprovidedwiththekit96determinationsStorageUsermanual1Closureplatemembrane2Sealedbags1Microelisastripplate12-8℃Standard:900pg/ml0.5ml×1bottle2-8℃Standarddiluent1.5ml×1bottle2-8℃HRP-Conjugatereagent6ml×1bottle2-8℃Samplediluent6ml×1bottle2-8℃ChromogenSolutionA6ml×1bottle2-8℃ChromogenSolutionB6ml×1bottle2-8℃StopSolution6ml×1bottle2-8℃washsolution30×20ml×1bottle2-8℃MATERIALSREQUIREDBUTNOTPROVIDEDlMicroplatereadercapableofmeasuringabsorbanceat450nm.lPrecisionpipettestodeliver2mlto1mlvolumes.l100mland1litergraduatedcylinders.lCalibratedadjustableprecisionpipettes,preferablywithdisposableplastictips.(Amanifoldmulti-channelpipetteisdesirableforlargeassays.)lAbsorbentpaper.l37°Cincubator.lDistilledordeionizedwater.lDataanalysisandgraphingsoftware.Graphpaper:linear(Cartesian),log-logorsemi-log,orlog-logitasdesired.lTubestopreparestandardorsampledilutions.STORAGECONDITIONSuWhenstoredat2°Cto8°Cunopenedreagentswillretainreactivityuntilexpirationdate.uDonotusereagentsbeyondthisdate.Openedreagentsmustbestoredat2°Cto8°C.uMicrotiterwellsmustbestoredat2°Cto8°C.Oncethefoilbaghasbeenopened,care首ldbetakentocloseittightlyagain.uOpenedkitsretainactivityfor8weeksifstoredasdescribedabove.REAGENTPREPARATIONBringallreagentstoroomtemperaturebeforeuseSPECIMENCOLLECTIONANDPREPARATIONSerum-Useaserumseparatortube(SST)andallowsamplestoclotfor30minutesbeforecentrifugationfor15minutesatapproximately1000xg.Removeserumandassayimmediatelyoraliquotandstoresamplesat-20°Cor-80°C.Plasma-CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor15minutesat1000xgat2-8°Cwithin30minutesofcollection.Storesamplesat-20°Cor-80°C.Avoidrepeatedfreeze-thawcycles.Cellculturefluidandotherbiologicalfluids-Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat-20°Cor-80°C.Avoidrepeatedfreeze-thawcyclesASSAYPROCEDUREuGeneralRemarkslAllreagentsandspecimensmustbeallowedtocometoroomtemperaturebeforeuse.Allreagentsmustbemixedwithoutfoaming.lOncethetesthasbeenstarted,allsteps首ldbecompletedwithoutinterruption.lUsenewdisposalplasticpipettetipsforeachstandard,controlorsampleinordertoavoidcrosscontamination.lAbsorbanceisafunctionoftheincubationtimeandtemperature.Beforestartingtheassay,itisrecommendedthatallreagentsareready,capsremoved,allneededwellssecuredinholder,etc.Thiswillensureequalelapsedtimeforeachpipettingstepwithoutinterruption.lAsageneralruletheenzymaticreactionislinearlyproportionaltotimeandtemperature.lDetermineabsorptionwithanELISAreaderat450nmagainst620nmasreference.Ifnoreferencewavelengthisavailable,readonlyat450nm.Iftheextinctionofthehigheststandardexceedsthemeasurementrangeofthephotometer,absorptionmustbemeasuredimmediatelyat405nmagainst620nmasreference.uAssayProcedure1.DiluteandaddsampletoStandard:set10StandardwellsontheELISAplatescoated,addStandard100μltothefirstandthesecondwell,thenaddStandarddilution50μltothefirstandthesecondwell,mix;takeout100μlformthefirstandthesecondwellthenaddittothethirdandtheforthwellseparately.thenaddStandarddilution50μltothethirdandtheforthwell,mix;thentakeout50μlfromthethirdandtheforthwelldiscard,add50μltothefifthandthesixthwell,thenaddStandarddilution50μltothefifthandthesixthwell,mix;takeout50μlfromthefifthandthesixthwellandaddtotheseventhandtheeighthwell,thenaddStandarddilution50μltotheseventhandtheeighthwell,mix;takeout50μlfromtheseventhandtheeighthwellandaddtotheninthandthetenthwell,addStandarddilution50μltotheninthandthetenthwell,mix,takeout50μlfromtheninthandthetenthwelldiscard(addSample50μltoeachwellafterDiluting,(density:600pg/ml,400pg/ml,200pg/ml,100pg/ml,50pg/ml).50pg/ml100pg/ml600pg/ml200pg/ml900pg/ml400pg/ml2.addsample:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.3.Incubate:AfterclosingplatewithClosureplatemembrane,incubatefor30minat37℃.4.Configurateliquid:30-foldwashsolutiondiluted30-foldwithdistilledwaterandreserve.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.CALCULATIONOFRESULTSlCalculatetheaverageabsorbancevaluesforeachsetofstandards,controlsandpatientsamples.lConstructastandardcurvebyplottingthemeanabsorbanceobtainedfromeachstandardagainstits.lconcentrationwithabsorbancevalueonthevertical(Y)axisandconcentrationonthehorizontal(X)axis.lUsingthemeanabsorbancevalueforeachsampledeterminethecorrespondingconcentrationfromthestandardcurve.lAutomatedmethod:TheresultsintheIFUhavebeencalculatedautomaticallyusinga4PL.l(4ParameterLogistics)curvefit.4ParameterLogisticsisthepreferredcalculationmethod.Otherdata.lreductionfunctionsmaygiveslightlydifferentresults.lTheconcentrationofthesamplescanbereaddirectlyfromthisstandardcurve.Sampleswith.lconcentrationshigherthanthatofthehigheststandardhavetobefurtherdiluted.Forthecalculationof.ltheconcentrationsthisdilutionfactorhastobetakenintoaccount.REFERENCESREF:Cat.-No.:/Kat.-Nr.:/No.-Cat.:/Cat.-No.:/N.Cat.:/N.CatLOT:Lot-No.:/Chargen-Bez.:/No.Lot:/Lot-No.:/LoteN.:/Lotton.::No.ofTests:/Kitgre:/Nb.deTests:/No.deDeterm.:/N.deTestes:/Quantitàdeitests::Keepawayfromheatordirectsunlight./VorHitzeunddirekterSonneneinstrahlungschützen./Garderàl’abridelachaleuretdetouteexpositionlumineuse./Manténgasealejadodelcalorolaluzsolardirecta./Manterlongedocalorouluzsolardirecta./Nonesporreairaggisolari.:Readinstructionsbeforeuse./Arbeitsanleitunglesen./Lirelafichetechniqueavantemploi./Lealasinstruccionesantesdeusar./Lerasinstruesantesdeusar./Leggereleistruzioniprimadell’uso.:Storeat:/Lagernbei:/Stockerà:/Almacenea:/Armazenara:/Conservarea:[详细]
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2018-09-22 10:00
产品样册
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Mouse (VLDL)ELISA Kit
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Mouseverylowdensitylipoprotein(VLDL)ELISAKitFORRESEARCHUSEONLY.NotforclinicaldiagnosisuseCATALOG#:DAG859INTRODUCTION?ThiskitallowsforthedeterminationofVLDLconcentrationsinMouseserum?Detectionofspecies:Mouse?Detectionmedium:serum,cellculturesupernates.?Assayrange:6.0μg/ml-160μg/mlPRINCIPLEOFTESTThekitassayMouseVLDLlevelinthesample,usePurifiedMouseVLDLantibodytocoatmicrotiterplatewells,makesolid-phaseantibody,thenaddVLDLtowells,CombinedVLDLantibodywhichWithHRPlabeled,becomeantibody-antigen-enzyme-antibodycomplex,afterwashingCompletely,AddTMBsubstratesolution,TMBsubstratebecomesbluecolorAtHRPenzyme-catalyzed,reactionisterminatedbytheadditionofasulphuricacidsolutionandthecolorchangeismeasuredspectrophotometricallyatawavelengthof450nm.TheconcentrationofMouseVLDLinthesamplesisthendeterminedbycomparingtheO.D.ofthesamplestothestandardcurve.Phone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG6342COMPOSITIONOFTHEKIT1washsolution20ml×1bottle7StopSolution6ml×1bottle2HRP-Conjugatereagent6ml×1bottle8Standard(320μg/ml)0.5ml×1bottle3Microelisastripplate12well×8strips9Standarddiluent1.5ml×1bottle4Samplediluent6ml×1bottle10Instruction15ChromogenSolutionA6ml×1bottle11Closureplatemembrane26ChromogenSolutionB6ml×1bottle12Sealedbags1STORAGECONDITIONS?Theunopenedkitshallbestoredat[2-8℃].?Foropenedkitcanbestoredat[2-8℃]forupto1month.Ifnotbeusedrecently,thestandard首ldbekeptin-20℃.WASHINGMETHOD?Manuallywashingmethod:shakeawaytheremainedliquidintheenzymeplates;placesomebibulouspapersonthetest-bed,andflaptheplatesontheupsidedownstrongly.Injectatleast0.35mlafter-dilutionwashingsolutionintothewell,andmarinate1~2minutes.Repeatthisprocessaccordingtoyourrequirements.?Automaticwashingmethod:ifthereisautomaticwashingmachine,it首ldonlybeusedinthetestwhenyouarequitefamiliarwithitsfunctionandperformance.SAMPLEPREPARATION1.extractassoonaspossibleafterSpecimencollection,andaccordingtotherelevantliterature,and首ldbeexperimentassoonaspossibleaftertheextraction.Ifitcan’t,specimencanbekeptin-20℃topreserve,Avoidrepeatedfreeze-thawcycles.Phone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG63432.Can’tdetectthesamplewhichcontainNaN3,becauseNaN3inhibitsHRPactive.ASSAYPROCEDUREStep1:Diluteandaddsample:DiluteOriginaldensityStandardasfollowtable:Step2:Setblankwellsseparately(blankcomparisonwellsdon’taddsampleandHRP-Conjugatereagent,othereachstepoperationissame).testingsamplewell.addSampledilution40μltotestingsamplewell,thenaddtestingsample10μl(samplefinaldilutionis5-fold),addsampletowells,don’ttouchthewellwallasfaraspossible,andGentlymix.Step3:Incubate:Coverwiththeadhesivestripprovided,incubatefor30minat37℃.Step4:Configurateliquid:Dilutewashsolution30-fold(or20-fold)withdistilledwater.Step5:Washing:Uncovertheadhesivestrip,discardliquid,Pipettewashingbuffertoeverywell,stillfor30sthendrain,repeat5times.Step6:Addenzyme:PipetteHRP-Conjugatereagent50μltoeachwell,exceptblankwell.Step7:Incubate:Operationwith3.Step8:Washing:Operationwith5.160μg/ml5Standard150μlOriginaldensityStandard+150μlStandarddiluent80μg/ml4Standard150μl5Standard+150μlStandarddiluent40μg/ml3Standard150μl4Standard+150μlStandarddiluent20μg/ml2Standard150μl3Standard+150μlStandarddiluent10μg/ml1Standard150μl2Standard+150μlStandarddiluentPhone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG6344Step9:Color:PipetteChromogenSolutionA50ulandChromogenSolutionBtoeachwell,avoidthelightpreservationfor15minat37℃Step10:Stopthereaction:PipetteStopSolution50μltoeachwell,Stopthereaction(thebluechangetoyellow).Step11:Calculate:takeblankwellaszero,Readabsorbanceat450nmafterPipetteingStopSolutionwithin15min.CALCULATIONOFRESULTTakethestandarddensityasthehorizontal,theODvalueforthevertical,drawthestandardcurveongraphpaper,FindoutthecorrespondingdensityaccordingtothesampleODvaluebytheSamplecurve,multipliedbythedilutionmultiple,orcalculatethestraightlineregressionequationofthestandardcurvewiththestandarddensityandtheODvalue,withthesampleODvalueintheequation,calculatethesampledensity,multipliedbythedilutionfactor,theresultisthesampleactualdensity.EXPIRATIONSixmonths[seelabelontheouterboxforthespecificdate].Phone:(612)379-2956Phone:(800)343-7475Fax:(612)656-4400Catalog#:DAG634TTENTION?Thekittakesoutfromtherefrigeration首ldbebalanced15-30minutesintheroomtemperature,ifthecoatedELISAplateshavenotbeenusedupafteropening,theplate首ldbestoredinsealedbag.?washingbufferwillCrystallizationseparation,itcanbeheatedthewaterhelpsdissolvewhendilute.Washingdoesnotaffecttheresult.?addSamplewithsamplerEachstep,Andproofreaditsaccuracyfrequently,avoidstheexperimentalerror.addsamplewithin5min,ifthenumberofsampleismuch,recommendtouseVolley.?ifthetestingmaterialcontentisexcessivelyhigher(ThesampleODisbiggerthanthefirststandardwell),pleasediluteSample(n-fold),Pleasediluenteandmultipliedbythedilutionfactor.(×n×5).?Closureplatemembraneonlylimitsthedisposableuse,toavoidcross-contamination.?Thesubstrate首ldevadethelighttobepreserved.?Pleaserefertotheuserinstructionstrictly,thetestresultdeterminationmusttakethemicrotiterplatereaderasastandard.?Thepreparationofsamplesandallthereagents首ldrefertoinfectivematerialprocess.?Donotmixreagentswiththosefromotherlots[详细]
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2018-10-31 10:00
产品样册
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人ELISA试剂盒,TPA ELISA Kit
- 96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原(TPA)ELISA试剂盒相关产品:96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原(TPA)ELISA试剂盒IL-10,小鼠白介素-10Elisa试剂盒磺胺二甲基恶唑,标准品JLJL200712人Elisa试剂盒,人ProteinCElisa试剂盒,HumanProteinCELISA试剂盒CAS号:108-69-0,3,5-二,98%人Elisa试剂盒,人CD30Elisa试剂盒,HumanClusterofdifferentiation30,CD30ELISA试剂盒CAS:893-36-7,盐酸-L-白氨酰-2-萘胺/L-亮氨酰-2-萘胺盐酸盐/L-白氨酰-β-萘胺盐酸盐/盐酸-L-亮氨酰-2-萘胺/L(+)-亮氨酰-2-萘基盐酸氨/L-Leucyl-2-naphthylamidehydrochloride,BR,98%,1克,避光,-20℃96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原(TPA)ELISA试剂盒Humanbacterialvaginosis,BVELISA试剂盒人(BV)kit说明书,细菌性阴道病Elisa试剂盒CXCR3ELISAKit,大鼠CXC趋化因子受体3Elisa检测试剂盒蒙花苷,标准品,含量测定,20mg,常温,避光PorcineapoproteinA1,apo-A1ELISA试剂盒猪(apo-A1)kit说明书,载脂蛋白A1Elisa试剂盒大鼠淋巴细胞因子ELISA试剂盒HumanhepatitisBvirusXinteractingprotein,HBXIPELISAKit人异常凝血酶原(APT)ELISA试剂盒HumanAbnormalprothrombin,APTELISA试剂盒草乌甲素,标准品,含量测定,50mg,常温,避光96孔elisa,48孔elisa,HumanTissuePolypeptideAntigen,TPAELISAKit,人组织多肽抗原(TPA)ELISA试剂盒GRP1957,营养肉汤,供一般细菌培养、转种和增菌用,250g小鼠生长激素释放多肽(GHRP)ELISA试剂盒HumanMotilin,MTLELISAKitCAS号:4767-3-7,2,2-双(羟甲基)丙酸,98%[详细]
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2018-10-23 10:31
产品样册
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[ELISA]CD8分子(CD8)ELISA Kit
- 羊CD8分子(CD8)试剂盒使用说明书本试剂盒仅供研究使用。检测范围:96T7IU/ml-240IU/ml试剂盒组成130倍浓缩洗涤液20ml×1瓶7终止液6ml×1瓶2酶标试剂6ml×1瓶8标准品(400IU/ml)0.5ml×1瓶3酶标包被板12孔×8条9标准品稀释液1.5ml×1瓶4样品稀释液6ml×1瓶10说明书1份5显色剂A液6ml×1瓶11封板膜2张6显色剂B液6ml×1/瓶12密封袋1个标本要求1.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。若不能马上进行试验,可将标本放于-20℃保存,但应避免反复冻融2.不能检测含NaN3的样品,因NaN3YZ辣根过氧化物酶的(HRP)活性。羊CD8分子ELISA试剂盒用于测定羊血清、血浆及相关液体样本中CD8分子(CD8)含量。操作步骤1.标准品的稀释:本试剂盒提供原倍标准品一支,用户可按照下列图表在小试管中进行稀释。200IU/ml5号标准品150μl的原倍标准品加入150μl标准品稀释液100IU/ml4号标准品150μl的5号标准品加入150μl标准品稀释液50IU/ml3号标准品150μl的4号标准品加入150μl标准品稀释液25IU/ml2号标准品150μl的3号标准品加入150μl标准品稀释液12.5IU/ml1号标准品150μl的2号标准品加入150μl标准品稀释液2.加样:分别设空白孔(空白对照孔不加样品及酶标试剂,其余各步操作相同)、标准孔、待测样品孔。在酶标包被板上标准品准确加样50μl,待测样品孔中先加样品稀释液40μl,然后再加待测样品10μl(样品Z终稀释度为5倍)。加样将样品加于酶标板孔底部,尽量不触及孔壁,轻轻晃动混匀。3.温育:用封板膜封板后置37℃温育30分钟。4.配液:将30倍浓缩洗涤液用蒸馏水30倍稀释后备用5.洗涤:小心揭掉封板膜,弃去液体,甩干,每孔加满洗涤液,静置30秒后弃去,如此重复5次,拍干。6.加酶:每孔加入酶标试剂50μl,空白孔除外。7.温育:操作同3。8.洗涤:操作同5。9.显色:每孔先加入显色剂A50μl,再加入显色剂B50μl,轻轻震荡混匀,37℃避光显色15分钟.10.终止:每孔加终止液50μl,终止反应(此时蓝色立转黄色)。11.测定:以空白空调零,450nm波长依序测量各孔的吸光度(OD值)。测定应在加终止液后15分钟以内进行。羊CD8分子ELISA试剂盒注意事项1.试剂盒从冷藏环境中取出应在室温平衡15-30分钟后方可使用,酶标包被板开封后如未用完,板条应装入密封袋中保存。2.浓洗涤液可能会有结晶析出,稀释时可在水浴中加温助溶,洗涤时不影响结果。3.各步加样均应使用加样器,并经常校对其准确性,以避免试验误差。一次加样时间**控制在5分钟内,如标本数量多,推荐使用排枪加样。4.请每次测定的同时做标准曲线,**做复孔。如标本中待测物质含量过高(样本OD值大于标准品孔**孔的OD值),请先用样品稀释液稀释一定倍数(n倍)后再测定,计算时请Z后乘以总稀释倍数(×n×5)。5.封板膜只限一次性使用,以避免交叉污染。6.底物请避光保存。7.严格按照说明书的操作进行,试验结果判定必须以酶标仪读数为准.8.所有样品,洗涤液和各种废弃物都应按传染物处理。9.本试剂不同批号组分不得混用。10.如与英文说明书有异,以英文说明书为准。保存条件及有效期1.试剂盒保存:;2-8℃。2.有效期:6个月羊CD8分子ELISA试剂盒应用双抗体夹心法测定标本中羊CD8分子(CD8)水平。用纯化的羊CD8分子(CD8)抗体包被微孔板,制成固相抗体,往包被单抗的微孔中依次加入CD8分子(CD8),再与HRP标记的CD8分子(CD8)抗体结合,形成抗体-抗原-酶标抗体复合物,经过彻底洗涤后加底物TMB显色。TMB在HRP酶的催化下转化成蓝色,并在酸的作用下转化成Z终的黄色。颜色的深浅和样品中的CD8分子(CD8)呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),通过标准曲线计算样品中羊CD8分子(CD8)浓度。[详细]
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2018-11-16 10:02
产品样册
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猪血纤蛋白原降解产物(FDP)ELISA试剂盒
- 猪血纤蛋白原降解产物(FDP)ELISA试剂盒[详细]
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2013-12-12 00:00
课件
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鲤鱼卵黄蛋白原(VTG)Elisa试剂盒使用说明书
- 本试剂盒用于测定鲤鱼血清、血浆及相关液体样本中卵黄蛋白原(VTG)含量。实验原理本试剂盒应用双抗体夹心法测定标本中鲤鱼卵黄蛋白原(VTG)水平。用纯化的鲤鱼卵黄蛋白原(VTG)抗体包被微孔板,制成固相抗体,往包被单抗的微孔中依次加入卵黄蛋白(VTG),再与HRP标记的卵黄蛋白原(VTG)抗体结合,形成抗体-抗原-酶标抗体复合物,经过彻底洗涤后加底物TMB显色。TMB在HRP酶的催化下转化成蓝色,并在酸的作用下转化成Z终的黄色。颜色的深浅和样品中的卵黄蛋白原(VTG)呈正相关。用酶标仪在450nm波长下测定吸光度(OD值),通过标准曲线计算样品中鲤鱼卵黄蛋白原(VTG)浓度。试剂盒组成130倍浓缩洗涤液20ml×1瓶7终止液6ml×1瓶2酶标试剂6ml×1瓶8标准品(1600μg/L)0.5ml×1瓶3酶标包被板12孔×8条9标准品稀释液1.5ml×1瓶4样品稀释液6ml×1瓶10说明书1份5显色剂A液6ml×1瓶11封板膜2张6显色剂B液6ml×1/瓶12密封袋1个标本要求1.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。若不能马上进行试验,可将标本放于-20℃保存,但应避免反复冻融2.不能检测含NaN3的样品,因NaN3YZ辣根过氧化物酶的(HRP)活性。操作步骤1.标准品的稀释:本试剂盒提供原倍标准品一支,用户可按照下列图表在小试管中进行稀释。800μg/L5号标准品150μl的原倍标准品加入150μl标准品稀释液400μg/L4号标准品150μl的5号标准品加入150μl标准品稀释液200μg/L3号标准品150μl的4号标准品加入150μl标准品稀释液100μg/L2号标准品150μl的3号标准品加入150μl标准品稀释液50μg/L1号标准品150μl的2号标准品加入150μl标准品稀释液2.加样:分别设空白孔(空白对照孔不加样品及酶标试剂,其余各步操作相同)、标准孔、待测样品孔。在酶标包被板上标准品准确加样50μl,待测样品孔中先加样品稀释液40μl,然后再加待测样品10μl(样品Z终稀释度为5倍)。加样将样品加于酶标板孔底部,尽量不触及孔壁,轻轻晃动混匀。3.温育:用封板膜封板后置37℃温育30分钟。4.配液:将30倍浓缩洗涤液用蒸馏水30倍稀释后备用5.洗涤:小心揭掉封板膜,弃去液体,甩干,每孔加满洗涤液,静置30秒后弃去,如此重复5次,拍干。6.加酶:每孔加入酶标试剂50μl,空白孔除外。7.温育:操作同3。8.洗涤:操作同5。9.显色:每孔先加入显色剂A50μl,再加入显色剂B50μl,轻轻震荡混匀,37℃避光显色15分钟.10.终止:每孔加终止液50μl,终止反应(此时蓝色立转黄色)。11.测定:以空白空调零,450nm波长依序测量各孔的吸光度(OD值)。测定应在加终止液后15分钟以内进行。计算以标准物的浓度为横坐标,OD值为纵坐标,在坐标纸上绘出标准曲线,根据样品的OD值由标准曲线查出相应的浓度;再乘以稀释倍数;或用标准物的浓度与OD值计算出标准曲线的直线回归方程式,将样品的OD值代入方程式,计算出样品浓度,再乘以稀释倍数,即为样品的实际浓度。[详细]
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2018-09-19 10:00
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