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AntibodiesOnline/IL 18 ELISA Kit (Interleukin 18)/ABIN2642443/10 x 96 tests188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
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AntibodiesOnline/IL 18 ELISA Kit (Interleukin 18)/ABIN2642443/10 x 96 tests
Antigen
Interleukin18(IL18)
  • Igif
  • Il-18
  • IL-18
  • IGIF
  • ChIL-18
  • IL-1g
  • IL1F4
  • IL18
  • interleukin18
  • interleukin18(interferon-gamma-inducingfactor)
  • interleukin-18
  • Il18
  • IL18
  • Il-18
Alternatives
HumanIL18ELISAKit
Kitswithalternativereactivityto:
32Human
26Rat(Rattus)
18Mouse(Murine)
17Pig(Porcine)
10Chicken
10Cow(Bovine)
10Rabbit
8Dog(Canine)
5Horse(Equine)
4Goat
4GuineaPig
2Monkey
2Wildboar(Susscrofa)
1Sheep(Ovine)
MethodType
SandwichELISA
DetectionRange
31.25-2000pg/mL
MinimumDetectionLimit
31.25pg/mL
Application
ELISA
Options
Bulkdiscount
Supplier
SupplierProductNo.
Independent Validation Badge
  • Successfullyvalidated(ELISA)
  • by AlamoLaboratoriesInc
  • No.#029598
  • Date02/09/2014
  •  
  • Readfullreport(PDF)
  • Clicktoverify
LotNumberU13095999
MethodvalidatedELISA
PositiveControlHumanserum
NegativeControlGoatserum
NotesSignalwasdetectedinpositivecontrolsampleandnotinnegativecontrolsample.
PrimaryAntibody
  • Antigen:Interleukin-18(IL-18)
  • Catalognumber:ABIN365215
  • Supplier:Logintosee
  • Suppliercatalognumber:Logintosee
  • Lotnumber:U13095999
Controls
  • Positivecontrol:Serumfromnormaladulthuman(specimenknowntocontainthetargetprotein).
  • Negativecontrol:Serumfromnormalgoat(specimensknowntonotcontainthetargetprotein).
  • Standardcurve:Serialtwo-folddilutionsfrom500pg/ml[500,250,125,62.5,31.3,15.6,7.8,0]weregeneratedfromthestandardprovidedinthekitusingstandard/samplediluentbuffer.
  • Spikecontrol:Standarddilutedinstandard/PBSdiluentbuffer[31.2and0].
Protocol
  • AllreagentsintheELISAkitwerebroughtuptoroomtemperature(RT)beforeuse.
  • 100µLofstandardorsamplewereaddedtowellsinELISAplatepre-coatedwithcaptureantibody.Allsamplesandstandardswereassayedintriplicate.
  • Theplatewascoveredwithsealer(providedinkit)andincubatedfor2hoursat37°C.UnboundmaterialwasaspiratedbutthewellswereNOTWashed.
  • 100µLofBiotin-Antibody(diluted1:100in“Biotin-AntibodyDiluent”)wasaddedtoeachwell.Platewascoveredwithsealer(providedinkit)andincubatedfor1hourat37°C.UnboundBiotin-Antibodywasremovedfromeachwellandplatewaswashedthreetimeswith350µLofwashbuffer(providedinthekit).Afterthelastwashtheplatewasinvertedagainstcleanabsorbentpapertoremoveanyremainingliquid.
  • 100µLofHRP-AvidinConjugate(diluted1:100in“HRP-AvidinDiluent”)wasaddedtoeachwell.Platewascoveredwithsealer(providedinkit)andincubatedfor1hourat37°C.
  • UnboundHRP-Avidinwasremovedbywashingfivetimeswith350µLofwashbuffer(providedinthekit).Afterthelastwashtheplatewasinvertedandblottedagainstcleanabsorbentpapertoremoveanyremainingliquid.
  • 90µLofTMBsubstratewasaddedtowellsandtheplatewascoveredwithanewplatesealer.Theplatewasgentlytappedtoensuremixingandincubatedfor30minat37°Cinthedark.
  • After30min,whenanapparentgradientappearedinthestandardwells,thereactionwasterminatedbyadding50µLofStopSolutiontoeachwell.
  • Theopticaldensity(ODvalue)ofeachwellwasreadusingamicroplatereadersetto450nm.
  • Thetriplicatereadingsforeachsamplewereaveragedandtheaveragezerostandardopticaldensitysubtractedtoyield‘correctedabsorbanceat450nm’.AstandardcurvewasgeneratedbyplottingthemeanODvalueforeachstandardontheX-axisagainsttheconcentrationontheY-axisusingExcel.Standardcurvewasgeneratedbyregressionanalysiswithfour-parameterlogistic.
  • Anequation(y=290.08x4-328.73x3+363.19x2+374.2x)wasderivedfromthestandardcurveandusedtocalculateInterleukin-18concentrationsinsamplesbasedontheirAverageAbsorbancevalues.
ExperimentalNotesNone

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