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当前位置： 首页 > 产品中心 > Other_kits > Diapharma/REAADS vWF Antigen/K034-001/Kit/96 tests
商品详细Diapharma/REAADS vWF Antigen/K034-001/Kit/96 tests
Diapharma/REAADS vWF Antigen/K034-001/Kit/96 tests
Diapharma/REAADS vWF Antigen/K034-001/Kit/96 tests
商品编号： K034-001
品牌： Diapharma
市场价： ¥0.00
美元价： 0.00
产地： 美国(厂家直采)
公司：
产品分类： 其它检测试剂盒
公司分类： Other_kits
联系Q Q： 3392242852
电话号码： 4000-520-616
电子邮箱： info@ebiomall.com
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商品介绍

Description:

TheREAADSvWFAntigen(vWF:Ag)testkitisanenzymelinkedimmunosorbentassayfordeterminingvonWillebrandFactorlevelsincitratedhumanplasma,expressedinrelativepercent(%)ofnormal.TheassayisintendedtobeusedasanaidinthediagnosisofvonWillebrandDiseaseinpatientswithbleedingdisorders,andtohelpdistinguishbetweenvWDandclassicHemophiliaA.REAADSvWF:AgTestKitwillaccuratelydetectantigenlevelsaslowas5%ofnormal.

Features:

  • ConvenientELISAprocedure
  • Objective,accurateandreproducIBLe
  • Reagentcompletekit
  • Totalincubationtime:40minutes

KitComposition:

Reagents

  • 12x8anti-humanVonWillebrandFactorantibodycoatedmicrowells.
  • 60mlSampleDiluent(blue-greensolution);containssodiumazide.
  • 3x0.5mllyophilizedReferencePlasma,withassaysheet.
  • 12mlanti-humanVWFHRPConjugate(redsolution).
  • 13mlSubstrate(TMBandH2O2).
  • 15mlStoppingSolution(0.36Nsulfuricacid).
  • 30mlWashConcentrate(33Xphosphatebufferedsalinewith0.01%Tween20).Note:turbiditymayappearinwashconcentratewhichwillnotaffectcomponentperformanceandshoulddisappearwhenworkingdilutionisprepared.

MaterialsRequiredbutnotSupplied

  • vWF:AgControlPlasma.Followmanufacturer’sinstructions,andstoreasrecommended.
  • Reagentgradewater(1L)topreparePBS/Tween20washsolution,toreconstituteReferencePlasma,andtozeroorblanktheplatereaderduringthefinalassaystep
  • Graduatedcylinders
  • Precisionpipettorscapableofdeliveringbetween5and1000µl,withappropriatetips
  • Miscellaneousglasswareappropriateforsmallvolumehandling3
  • Flaskorbottle,1liter
  • Washbottles,preferablywiththetippartiallycutbacktoprovideawidestream,oranautomatedorsemi-automatedwashingsystem
  • Disposablegloves,powder-freerecommended
  • PlatereADIngspectrophotometercapableofreadingabsorbanceat450nm(witha650nmreferenceifavailable)
  • Multichannelpipettorscapableofdeliveringto8wellssimultaneously
  • Microdilutiontubesforpatientsamplepreparation

PrincipleandProcedure:

Principle

TheREAADSvonWillebrandFactorAntigen(vWF:Ag)TestKitisanenzymelinkedimmunosorbentassayfordeterminingvonWillebrandFactorlevelsinhumanplasma,expressedinrelativepercent(%)ofnormal.TheassayisintendedtobeusedasanaidinthediagnosisofvonWillebrandDiseaseinpatientswithbleedingdisorders,andtohelpdistinguishbetweenvWDandclassicHemophiliaA.REAADSvWF:Agtestkitwillaccuratelydetectantigenlevelsaslowas5%ofnormal.

Procedure

DilutedcitratedpatientplasmaandcontrolsareincubatedinmicrowellscoatedwithcaptureantibodyspecificforhumanvWF,allowingpatientvWFtobindtothesurface.Followinganincubationperiod,thewellsarewashed,andahorseradishperoxidase(HRP)conjugatedanti-humanvWFdetectionantibodyisadded.Afterincubation,thewellsarewashed,substrateisadded,andcolordevelopmentismeasuredinaspectrophotometerat450nmfollowingtheadditionofastopsolution.PatientvWF:Aglevelsaredeterminedfromasix-pointcurvepreparedfromthereferenceplasmaprovidedinthekit.Totalincubationtimeis40minutesatroomtemperature.

AssayProcedure:

1.Removeanymicrowellstripsthatwillnotbeusedfromtheframeandstoretheminthebagprovided.

2.Assayeachreferenceplasmadilutioninduplicate.Duplicatedeterminationsarealsorecommendedforpatientandcontrolsamples.Onewellshouldberunasareagentblank;samplediluentwithoutplasmaisaddedtothewellasexplainedinstep6ofthissection.Thiswellistreatedthesameasapatientsampleinsubsequentassaysteps.Awaterblankwellshouldbeincludedwitheachplate;itistoremainemptyuntil200μLofreagentgradewaterisaddedatthecompletionoftheassay,immediatelypriortoreadingtheplate.Thewaterblankwellistobeusedtozerotheplatereader.

3.UsingtheReferencePlasmaprovidedwiththekit,preparesixreferenceplasmadilutionsasdescribedbelow:

VolumeReferencePlasmaVolumeSampleDiluent*ReferenceLevel(%)
30μl+500μl==150
20μl+500μl=100
15μl+500μl=75
10μl+500μl=50
10μl+1000μl=25
10μl+2000μl=12.5
**10μl+**4000μl=6.25
*Referencelevelvaluetobeusedforconstructingreferencecurveonly.

**MakeoneadditionaldilutioniftheassayedvalueoftheReferencePlasmais≥150%.

 

4.Preparea1:26dilutionofeachpatientsampleandcontrolplasmaselectedforuseinSampleDiluent(blue-greensolution);e.g.20μlsampleaddedto500μlSampleDiluent.Mixthoroughly.

5.Add100μlofthedilutions(referenceplasmasx6,patientsamplesandcontrols)totheappropriatemicrowells.6.Add100μlofSampleDiluenttothereagentblankwell.Leavethewaterblankwellempty.

7.Incubate15minutesatroomtemperature.Aftertheincubationiscomplete,carefullyinvertthemicrowellsanddumpthefluid.Donotallowsamplestocontaminateothermicrowells.

8.Wash4timeswithworkingwashsolution(PBS/Tween20).Eachwellshouldbefilledwithwashsolutionperwash.Washsolutionintheemptywellintendedtoserveasawaterblankwillnotinterferewiththeprocedure.Invertmicrowellsbetweeneachwashtoemptyfluid.Useasnappingmotionofthewristtoshaketheliquidfromthewells.Theframemustbesqueezedatthecenteronthetopandbottomtoretainmicrowellmodulesduringwashing.Blotonabsorbentpapertoremoveresidualwashfluid.Donotallowwellstodryoutbetweensteps.

9.Add100μlConjugate(red)toeachwell(exceptthewaterblankwell).

10.Incubatefor15minutesatroomtemperature.Aftertheincubationiscomplete,carefullyinvertthemicrowellsanddumptheconjugatesolution.

11.Wash4timeswithworkingwashsolution(PBS/Tween20)asinstep8.Washsolutioninthewaterblankwellwillnotinterferewiththeprocedure.Useasnappingmotiontodraintheliquid,andblotonabsorbentpaperafterthefinalwash.Donotallowthewellstodryout.

12.Add100μlSubstratetoeachwell(exceptforthewaterblankwell)andincubatefor10minutesatroomtemperature.Addthesubstratetothewellsatasteadyrate.Bluecolorwilldevelopinwellswithpositivesamples.

13.Add100μlStoppingSolution(0.36Nsulfuricacid)toeachwell(exceptforthewaterblankwell)tostoptheenzymereaction.BesuretoaddStoppingSolutiontothewellsinthesameorderandatthesamerateastheSubstrateSolutionwasadded.Bluesubstratewillturnyellowandcolorlesssubstratewillremaincolorless.DonotaddStoppingSolutiontothewaterblankwell.Instead,add200μlreagentgradewatertothewaterblankwell.Blankorzerotheplatereaderagainstthewaterblankwell.ReadtheO.D.ofeachwellat450nm,againsta650nmreferencefilter(ifavailable).Forbestresults,theO.D.valuesshouldbemeasuredwithin30minutesaftertheadditionofStoppingSolution.

Performance:

ClinicalPerformance

TheclinicalperformancewasdeterminedbytestinghealthyblooddonorsandvonWillebrandDiseaseplasmasampleswithREAADSvWF:AgTestKitandwithawellestablished,commerciallyavailablevonWillebrandFactorAntigenRocketEIAmethod.Theresultscorrelatedwell,andwereshowntobestatisticallysimilarbysinglefactorAnova.

REAADSRocketEIA
Healthy–Mean106%103%
Healthy–Range47–197%47–202%
vWFsamples–Mean38.8%37.4%
vWFsamples–Range26–59%24–50%
Correlation(r)=0.962;Pvalue=0.739

TechnicalPerformance

Intra-assayprecisionofREAADSvWF:Agtestkitis3.6%,whileinter-assayprecisionis5.0%.Linearity,expressedasthecoefficientofdetermination(r2)is0.996,withameanaccuracyof103.6%.TheREAADSvWF:Agtestkitisarapid,convenient,highlyaccurateandprecisemethodforthequantitativedeterminationofvWF:Aglevels.

Background:

vonWillebrandFactor(vWF),aprocoagulantprotein,playstwoimportantrolesinhemostasis:

  1. mediatesplateletadhesiontositesofvascularinjury
  2. protectsfactorVIIIfromproteolyticcleavageincirculation.InvonWillebrandDisease(vWD),whichiscausedbyeitherquantitative(TypeI)orqualitative(TypeII)deficienciesinvWF,abnormalbleedingmayresultduetoimpairedplateletfunctionandclottingfactorinhibition.

vWDisthemostcommoninheritedbleedingdisorder,andisclinicallycharacterizedbyeasybruisingorprolongedbleedingfrommucosalsurfaces.Whileapproximately80%ofvWDpatientshaveTypeIdeficiency,bothquantitative(antigenic)andqualitative(functional)assaysmayberequiredforalaboratorydiagnosisofvWD.

vonWillebrandFactorAntigen(vWF:AgorFactorVIII-relatedprotein)isaplasmaproteinfoundincirculationcombinedbynon-covalentinteractionswithFactorVIII(FVIII:C),apro-coagulantproteinalsoknownastheanti-hemophilicfactor.Thesetwoproteinsshowdistinctbiochemicalandfunctionalpropertiesaswellasdifferentantigenicdeterminants;theirplasmalevelsmayvaryindependentlyofeachother.DeficiencyofFVIIIcausesclassichemophiliawhiledeficiencyofvWFcausesvonWillebranddisease.MostofvWF:Agissynthesizedandstoredbyendothelialcellswhile15-20%issynthesizedbymegakaryocytesandstoredincirculatingplatelets.AvWF:Agunithasamolecularweightofabout250kDandtendstopolymerizeincirculation,withmultimersranginginsizefrom850kDtoaslargeas15×106D.
vWF:Agplaysaveryimportantroleinhemostasis;itprotectsFVIIIfromproteolyticcleavageincirculationandhelpsplateletstoaggregateortoadheretositesofvasculardamage.Theinvivohalf-lifeofFVIII:CwithoutvWF:Agisshortenedfrom10-12hourstoafewminutes.Thesetwomechanismspreventbleeding.VonWillebranddiseaseischaracterizedbyaninheriteddeficiencyofvWF.AdecreasedvWFactivityinplasmacanbetheresultoflowconcentrations(quantitativeortypeIdefect)oradeficientfunctionofvWF(qualitativeortypeIIdefect). vonWillebranddiseaseisthemostcommoninheritedbleedingdisordercharacterizedbyeasybruisingandprolongedbleedingfrommucosalsurfaces.TheprevalenceofvonWillebranddiseasehasbeenestimatedtobe1-3%ofthegeneralpopulation.Approximately80%ofvonWillebranddiseasepatientshaveatypeIdeficiency.
ThelaboratorydiagnosisofvonWillebranddiseasemayrequirebothquantitativeandqualitative(functional)determinations.QuantitativedeterminationsarebasedonimmunologictechniquessuchasradialimmunodiffusioningelandLaurellrocketimmunoelectrophoresis.ELISAprocedures9appliedtomeasurevWF:Agarelesslaborintensiveandofferseveraladvantagesincludingmoreobjective,accurateandreproducibleresults.Inaddition,ELISAallowsautomationwithcommonlyavailablelaboratoryinstruments.
品牌介绍
Diapharma使命宣言位于俄亥俄州西切斯特的Difarma Group，Inc.在诊断和研究领域销售止血、血栓形成、血小板功能测试、仪器和凋亡产品，并提供强大的技术能力和经验，以确保满足或超过客户的期望。地黄止血显色凝块酶联免疫吸附试验试剂盒历史1997年1月1日，由俄亥俄州富兰克林市的Pharmacia Hepar，Inc.成立，最初是Chromogenix基质和分析的独家美国和加拿大经销商。四分之一个多世纪前，Chromogenix开发了第一个显色底物技术，其前身是Kabi Diagnostica。卡比后来与法玛西亚合并。希帕玛目前的一些员工在法玛西亚肝素制造厂的显色部门工作。1998年，夏帕玛搬到了俄亥俄州的西切斯特，至今仍在那里。多年来，迪法玛扩大了其产品线，包括各种止血、细胞死亡、血小板功能、生态毒理学、化验、试剂、抗体和高级制造商的仪器。2017年，夏帕玛庆祝了20年的成功
公司介绍
Diapharma使命宣言位于俄亥俄州西切斯特的Difarma Group，Inc.在诊断和研究领域销售止血、血栓形成、血小板功能测试、仪器和凋亡产品，并提供强大的技术能力和经验，以确保满足或超过客户的期望。地黄止血显色凝块酶联免疫吸附试验试剂盒历史1997年1月1日，由俄亥俄州富兰克林市的Pharmacia Hepar，Inc.成立，最初是Chromogenix基质和分析的独家美国和加拿大经销商
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