Everest Biotech/Human ADAMTS4 ELISA Kit 1 plate/2905920015/1 plate,Everest Biotech,,Everest Biotech,everestbiotech,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：Everest Biotech/Human ADAMTS4 ELISA Kit 1 plate/2905920015/1 plate

产品编号：2905920015

市场价：¥4461.60

场地：美国(厂家直采)

产品分类：试剂盒>ELISA试剂盒>夹心法ELISA>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：$429.00

品牌： Everest Biotech

公司：Everest Biotech

公司分类：

商品介绍

ForquantitativedetectionofhumanADAMTS4incellculturesupernates,serumandplasma(heparin,EDTA).

TypicalDataObtainedfromHumanADAMTS4

(TMBreactionincubateat37°Cfor25min)

Concentration(pg/ml)	0.0	625	1,250	2,500	5,000	10,000	20,000	40,000

O.D	0.046	0.110	0.159	0.293	0.478	0.821	1.386	1.833

TypicalHumanADAMTS4ELISAKitStandardCurve

ThisstandardcurvewasgeneratedatEtonfordemonstrationpurposeonly.Astandardcurvemustberunwitheachassay.

Range625pg/ml-40,000pg/ml

Sensitivity<20pg/ml

SpecificityNaturalandrecombinanthumanADAMTS4

Cross-reactivityNodetectablecross-reactivitywithotherrelevantproteins

Storage

Storeat4°Cfor6months,at-20°Cfor12months.Avoidmultiplefreeze-thawcycles(Shippedwithwetice.)

Precision

Intra-AssayPrecision(Precisionwithinanassay)Threesamplesofknownconcentrationweretestedononeplatetoassessintra-assayprecision.

Inter-AssayPrecision(Precisionbetweenassays)Threesamplesofknownconcentrationweretestedinseparateassaystoassessinter-assayprecision.

	Intra-AssayPrecision			Inter-AssayPrecision
Sample	1	2	3	1	2	3
n	16	16	16	24	24	24
Mean(ng/ml)	5.9	12	21	6.5	14	23
Standarddeviation	0.248	0.612	1.2	0.377	0.882	1.66
CV(%)	4.2	5.1	5.7	5.8	6.3	7.2

Principle

Eton’shumanADAMTS4ELISAKitwasbasedonstandardsandwichenzyme-linkedimmune-sorbentassaytechnology.AmonoclonalantibodyfrommousespecificforADAMTS4hasbeenprecoatedonto96-wellplates.Standards(CHO,F213-C685)andtestsamplesareaddedtothewells,abiotinylateddetectionpolyclonalantibodyfromgoatspecificforADAMTS4isaddedsubsequentlyandthenfollowedbywashingwithPBSorTBSbuffer.Avidin-Biotin-PeroxidaseComplexwasaddedandunboundconjugateswerewashedawaywithPBSorTBSbuffer.HRPsubstrateTMBwasusedtovisualizeHRPenzymaticreaction.TMBwascatalyzedbyHRPtoproduceabluecolorproductthatchangedintoyellowafteraddingacidicstopsolution.ThedensityofyellowisproportionaltothehumanADAMTS4amountofsamplecapturedinplate.

KitComponents

Description	Quantity
96-wellplateprecoatedwithanti-humanADAMTS4antibody	1
LyophilizedrecombinanthumanADAMTS4standard	40ng/tube×2
Biotinylatedanti-humanADAMTS4antibody	130μl(dilution1:100)
Avidin-Biotin-PeroxidaseComplex(ABC)	130μl(dilution1:100)
Samplediluentbuffer	30ml
Antibodydiluentbuffer	12ml
ABCdiluentbuffer	12ml
TMBcolordevelopingagent	10ml
TMBstopsolution	10ml

MaterialRequiredButNotProvided

1.Microplatereaderinstandardsize.

2.Automatedplatewasher.

3.AdjustablePipettesandpipettetips.Multichannelpipettesarerecommendedintheconditionoflargeamountofsamplesinthedetection.

4.CleantubesandEppendorftubes.

5.Washingbuffer(neutralPBSorTBS).

Preparationof0.01MTBS:Add1.2gTris,8.5gNacl;450μlofpurifiedaceticacidor700μlofconcentratedhydrochloricacidto1000mlH2OandadjustpHto7.2-7.6.Finally,adjustthetotalvolumeto1L.

Preparationof0.01MPBS:Add8.5gsodiumchloride,1.4gNa2HPO4and0.2gNaH2PO4to1000mldistilledwaterandadjustpHto7.2-7.6.Finally,adjustthetotalvolumeto1L.

NoticeforApplicationofKit

1.Toinspectthevalidityofexperimentoperationandtheappropriatenessofsampledilutionproportion,pilotexperimentusingstandardsandasmallnumberofsamplesisrecommended.

2.TheTMBColorDevelopingagentiscolorlessandtransparentbeforeusing,contactusfreelyifitisnotthecase.

3.BeforeusingtheKit,spintubesandbringdownallcomponentstothebottomoftubes.

4.Duplicatewellassayisrecommendedforbothstandardandsampletesting.

5.Don’tlet96-wellplatedry,fordryplatewillinactivateactivecomponentsonplate.

6.Don’treusetipsandtubestoavoidcrosscontamination.

7.Avoidusingthereagentsfromdifferentbatchestogether.

8.Inordertoavoidmarginaleffectofplateincubationduetotemperaturedifference(reactionmaybestrongerinthemarginalwells),itissuggestedthatthedilutedABCandTMBsolutionwillbepre-warmedin37℃for30minbeforeusing.

Preparation

1.SamplePreparationandStorage

Storesamplestobeassayedwithin24hoursat2-8°C.Forlong-termstorage,aliquotandfreezesamplesat-20°C.Avoidrepeatedfreeze-thawcycles.

Cellculturesupernates:Removeparticulatesbycentrifugation,assayimmediatelyoraliquotandstoresamplesat

-20°C.

Serum:Allowtheserumtoclotinaserumseparatortube(about4hours)atroomtemperature.Centrifugeatapproximately1000Xgfor15min.Analyzetheserumimmediatelyoraliquotandstorefrozenat-70°C.

Plasma:CollectplasmausingheparinorEDTAasananticoagulant.Centrifugefor15minat1000xgwithin30minofcollection.Analyzeimmediatelyoraliquotandstorefrozenat-70°C.Citrateisnotrecommendedastheanticoagulant.

2.SampleDilutionGuideline

Theuserneedstoestimatetheconcentrationofthetargetproteininthesampleandselectaproperdilutionfactorsothatthedilutedtargetproteinconcentrationfallsnearthemiddleofthelinearregimeinthestandardcurve.Dilutethesampleusingtheprovideddiluentbuffer.Thefollowingisaguidelineforsampledilution.Severaltrialsmaybenecessaryinpractice.Thesamplemustbewellmixedwiththediluentsbuffer.

Hightargetproteinconcentration(400-4000ng/ml).Theworkingdilutionis1:100.i.e.Add3μlsampleinto297μlsamplediluentbuffer.

Mediumtargetproteinconcentration(40-400ng/ml).Theworkingdilutionis1:10.i.e.Add25μlsampleinto225μlsamplediluentbuffer.

Lowtargetproteinconcentration(625-40,000pg/ml).Theworkingdilutionis1:2.i.e.Add100μlsampleto100μlsamplediluentbuffer.

VeryLowtargetproteinconcentration(≤625pg/ml).Nodilutionnecessary,ortheworkingdilutionis1:2.

3.ReagentPreparationandStorage

A.ReconstitutionofthehumanADAMTS4standard:ADAMTS4standardsolutionshouldbepreparednomorethan2hourspriortotheexperiment.TwotubesofADAMTS4standard(40ngpertube)areincludedineachkit.Useonetubeforeachexperiment.

a.40,000pg/mlofhumanADAMTS4standardsolution:Add1mlsamplediluentbufferintoonetube,keepthetubeatroomtemperaturefor10minandmixthoroughly.

b.20,000pg/ml→625pg/mlofhumanADAMTS4standardsolutions:Label6Eppendorftubeswith20,000pg/ml,10,000pg/ml,5,000pg/ml,2,500pg/ml,1,250pg/ml,625pg/mlrespectively.Aliquot0.3ml

ofthesamplediluentbufferintoeachtube.Add0.3mloftheabove40,000pg/mlADAMTS4standardsolutioninto1sttubeandmix.Transfer0.3mlfrom1sttubeto2ndtubeandmix.Transfer0.3mlfrom2ndtubeto3rdtubeandmix,andsoon.

Note:Thestandardsolutionsarebestusedwithin2hours.The40ng/mlstandardsolutionshouldbestoredat4°Cforupto12hours,orat-20°Cforupto48hours.Avoidrepeatedfreeze-thawcycles.

B.Preparationofbiotinylatedanti-humanADAMTS4antibodyworkingsolution:Thesolutionshouldbepreparednomorethan2hourspriortotheexperiment.

a.Thetotalvolumeshouldbe:0.1ml/wellx(thenumberofwells).(Allowing0.1-0.2mlmorethantotalvolume)

b.Biotinylatedanti-humanADAMTS4antibodyshouldbedilutedin1:100withtheantibodydiluentbufferandmixedthoroughly.(i.e.Add1μlBiotinylatedanti-humanADAMTS4antibodyto99μlantibodydiluentbuffer.)

C.PreparationofAvidin-Biotin-PeroxidaseComplex(ABC)workingsolution:Thesolutionshouldbepreparednomorethan1hourpriortotheexperiment.

a.Thetotalvolumeshouldbe:0.1ml/wellx(thenumberofwells).(Allowing0.1-0.2mlmorethantotalvolume)

b.Avidin-Biotin-PeroxidaseComplex(ABC)shouldbedilutedin1:100withtheABCdilutionbufferandmixedthoroughly.(i.e.Add1μlABCto99μlABCdiluentbuffer.)

AssayProcedure

TheABCworkingsolutionandTMBcolordevelopingagentmustbekeptwarmat37°Cfor30minbeforeuse.Whendilutingsamplesandreagents,theymustbemixedcompletelyandevenly.StandardADAMTS4detectioncurveshouldbepreparedforeachexperiment.TheuserwilldecidesampledilutionfoldbycrudeestimationofADAMTS4amountinsamples.

品牌介绍

Everest Biotech公司成立于2000年，由英国牛津大学一批生物化学家创建。公司科学家团队具有丰富的抗体纯化及蛋白组学专业知识，雄厚的专业背景赋予了公司能为全球科研人员提供高质量、创新性抗体产品的能力。目前，Everest Biotech可提供3000余种抗体，涉及肿瘤、自身免疫、过敏、心血管疾病、传染病、代谢、神经科学等领域。所有抗体均由Everest Biotech自主研发生产、全部经过肽亲和纯化以保证最佳的特异性和纯度、经过免疫原肽的ELISA和WB实验验证; 由PhD科学家团队提供全方位技术支持服务。Everest Biotech为众多国际知名抗体公司提供OEM服务。