产品资料

Antibodies-Online/Thrombospondin 1 ELISA Kit (THBS1)/ABIN2643294/5 x 96 tests

Antigen

TMTSP
UNQ3010
THBS
THBS-1
TSP
TSP-1
TSP1
CRISP-2
CT36
GAPDL5
TPX1
Thbs-1
tbsp1
thrombospondin-1
thbs
thbs-1
tsp
tsp-1
tsp1
Tsp1
THBS1
Tsp-1
wu:fc48c03
si:dkey-11e23.1
4833423O18Rik
Tmtsp
thrombospondin,typeI,domaincontaining1
thrombospondin1
cysteine-richsecretoryprotein2
thrombospondin,typeI,domain1
THSD1
THBS1
CRISP2
Thsd1
Thbs1
thbs1
LOC100230636

Alternatives

HumanThrombospondin1ELISAKit

MethodType

SandwichELISA

DetectionRange

7.8-500ng/mL

MinimumDetectionLimit

7.8ng/mL

Application

ELISA

Options

Supplier

SupplierProductNo.

	Successfullyvalidated(ELISA) by AlamoLaboratoriesInc No.#029601 Date02/09/2014 Readfullreport(PDF) Clicktoverify
LotNumber	T30095909
Methodvalidated	ELISA
PositiveControl	Humanserum
NegativeControl	Goatserum
Notes	SignalwasdetectedinpositivecontrolsampleandnegligIBLeamountweredetectedinnegativecontrolsample.

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PrimaryAntibody	Antigen:Humanthrombospondin1(TSP-1) Catalognumber:ABIN366828 Supplier:Logintosee Suppliecatalognumber:CSB-E08763h Lotnumber:T30095909
Controls	Positivecontrol:Serumfromnormaladulthuman(specimenknowntocontainthetargetprotein). Negativecontrol:Serumfromnormalgoat(specimensknowntonotcontainthetargetprotein). Standardcurve:Serialtwo-folddilutionsfrom500ng/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=78.721x4-235.57x3+279.75x2+48.209x)wasderivedfromthestandardcurveandusedtocalculateThrombospondin-1concentrationsinsamplesbasedontheirAverageAbsorbancevalues.
ExperimentalNotes	None

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Antibodies-Online/Thrombospondin 1 ELISA Kit (THBS1)/ABIN2643294/5 x 96 tests

FullMethods