货号：	MJ-R30986
样本：	大鼠血清、组织、尿液等
标记物：	检测指标
适应物种：	Rat
应用：	科研
检测方法：	双抗夹心
检测限：	见说明
供应商：	名劲生物
数量：	现货
规格：	96T/48T

Sample collectionandstorages

Serum -Useaserumseparatortubeandallowsamplestoclotfor30minutesbeforecentrifugationfor10minutesatapproximately3000×g.Removeserumandassayimmediatelyoraliquotandstoresamplesat-20℃or-80℃.Avoidrepeatedfreeze-thawcycles

Plasma -CollectplasmausingEDTAorheparinasananticoagulant.Centrifugesamplesfor30 minutesat3000×gat2-8℃within30minutesofcollection.Storesamplesat-20℃or-80℃.Avoidrepeatedfreeze-thawcycles.

Cellculturesupernatesandotherbiologicalfluids - Removeparticulatesbycentrifugationandassayimmediatelyoraliquotandstoresamplesat-20℃or-80℃.Avoidrepeatedfreeze-thawcycles.

Note: Thesamplesshoulebecentrifugateddequatelyandnohemolysisorgranulewasallowed.

Materialsrequiredbutnotsupplied
1. Standardmicroplatereader(450nm)
2. PrecisionpipettesandDisposablepipettetips.
3. 37℃incubator

Reagentpreparation
20×washsolution:DilutewithDistilledordeionizedwater 1:20.

Assayprocedure
1. Prepareall reagents before starting assay procedure. It is recommended that all Standards andSamples beadded induplicate totheMicroELISA Stripplate.
2. Addstandard:SetStandardwells,testingsamplewells.Addstandard50μltostandardwell.
3. AddSample:Addtestingsample 10μlthenaddSampleDiluent40μltotestingsamplewell;Blankwell doesn’taddanyting.
4. Add 100μl of HRP-conjugatereagent toeachwell, coverwithan adhesivestrip and incubate for 60minutes at 37°C. 
5. Aspirateeachwellandwash,repeatingtheprocessfour timesforatotaloffivewashes. WashbyfillingeachwellwithWashSolution (400μl)usingasquirtbottle,manifold dispenser orautowasher.Completeremovalofliquidateachstepisessentialtogoodperformance.Afterthelastwash,removeanyremainingWashSolution byaspiratingor decanting.Inverttheplateandblotitagainstcleanpapertowels.
6. AddchromogensolutionA50μlandchromogensolutionB50μltoeachwell. Gentlymixandincubatefor15minutesat37°C.

Protectfromlight.
7. Add50μlStopSolutiontoeachwell.Thecolorinthewellsshouldchangefromblueto yellow.Ifthecolorinthewellsisgreenorthecolorchangedoesnot
appearuniform, gentlytaptheplatetoensurethoroughmixing.
8. Read the Optical Density (O.D.) at 450 nm using amicrotiter plate reader within 15 minutes.

Calculationofresults

• Thisstandardcurveisusedtodeterminetheamountinanunknownsample.ThestandardcurveisgeneratedbyplottingtheaverageO.D.(450nm)obtainedforeachofthesixstandardconcentrationsonthevertical(Y)axisversusthecorrespondingconcentrationonthehorizontal(X)axis.
• First,calculatethemeanO.D.valueforeachstandardandsample.AllO.D.values,aresubtractedbythemeanvalueofthezerostandardbeforeresultinterpretation.Constructthestandardcurveusinggraphpaperorstatisticalsoftware.
• Todeterminetheamountineachsample,firstlocatetheO.D.valueontheY-axisandextendahorizontallinetothestandardcurve.Atthepointofintersection,drawaverticallinetotheX-axisandreadthecorrespondingconcentration.
• Anyvariationinoperator,pipettingandwashingtechnique,incubationtimeortemperature,andkitagecancausevariationinresult.Eachusershouldobtaintheirownstandardcurve.
• Thesensitivitybythisassayis 1.0pg/ml
• Standardcurve 

 Storage： 2-8℃.
validity：sixmonths.

FORRESEARCHUSEONLY;NOTFORTHERAPEUTICORDIAGNOSTICAPPLICATIONS!PLEASEREADTHROUGHENTIREPROCEDUREBEFOREBEGINNING!