Everest Biotech/Human Azurocidin ELISA Kit 1 plate/2904450015/1 plate,Everest Biotech,,Everest Biotech,everestbiotech,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：Everest Biotech/Human Azurocidin ELISA Kit 1 plate/2904450015/1 plate

产品编号：2904450015

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

ForquantitativedetectionofhumanAzurocidinincellculturesupernates,serumandplasma(heparin,EDTA).

TypicalDataObtainedfromHumanAzurocidin

(TMBreactionincubateat37°Cfor25min)

Concentration(pg/ml)	0.0	62.5	125	250	500	1000	2000	4000

O.D	0.015	0.046	0.070	0.121	0.293	0.650	1.514	2.260

TypicalHumanAzurocidinELISAKitStandardCurve

ThisstandardcurvewasgeneratedatEtonfordemonstrationpurposeonly.Astandardcurvemustberunwitheachassay.

Range62.5pg/ml-4000pg/ml

Sensitivity<10pg/ml

SpecificityNaturalandrecombinanthumanAzurocidin

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(pg/ml)	675	1847	2816	873	2020	3174

StandardDeviation	28.35	94.2	149.2	48.9	129.3	238.1

CV(%)	4.2	5.1	5.3	5.6	6.4	7.5

Principle

Eton’shumanAzurocidinELISAKitwasbasedonstandardsandwichenzyme-linkedimmune-sorbentassaytechnology.AmonoclonalantibodyfrommousespecificforAzurocidinhasbeenprecoatedonto96-wellplates.Standards(NSO,I27-P248)andtestsamplesareaddedtothewells,abiotinylateddetectionpolyclonalantibodyfromgoatspecificforAzurocidinisaddedsubsequentlyandthenfollowedbywashingwithPBSorTBSbuffer.Avidin-Biotin-PeroxidaseComplexwasaddedandunboundconjugateswerewashedawaywithPBSorTBSbuffer.HRPsubstrateTMBwasusedtovisualizeHRPenzymaticreaction.TMBwascatalyzedbyHRPtoproduceabluecolorproductthatchangedintoyellowafteraddingacidicstopsolution.ThedensityofyellowisproportionaltothehumanAzurocidinamountofsamplecapturedinplate.

KitComponents

Description	Quantity

96-wellplateprecoatedwithanti-humanAzurocidinantibody	1

LyophilizedrecombinanthumanAzurocidinstandard	10ng/tube×2

Biotinylatedanti-humanAzurocidinantibody	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.Analyzetheserumimmediatelyoraliquotandstoresamplesat-20°C.

Plasma:CollectplasmausingheparinorEDTAasananticoagulant.Centrifugefor15minat1500xgwithin30minofcollection.Assayimmediatelyoraliquotandstoresamplesat-20°C.

2.SampleDilutionGuideline

Theuserneedstoestimatetheconcentrationofthetargetproteininthesampleandselectaproperdilutionfactorsothatthedilutedtargetproteinconcentrationfallsnearthemiddleofthelinearregimeinthestandardcurve.Dilutethesampleusingtheprovideddiluentbuffer.Thefollowingisaguidelineforsampledilution.Severaltrialsmaybenecessaryinpractice.Thesamplemustbewellmixedwiththediluentsbuffer.

Hightargetproteinconcentration(40-400ng/ml).Theworkingdilutionis1:100.i.e.Add3μlsampleinto297μlsamplediluentbuffer.

Mediumtargetproteinconcentration(4-40ng/ml).Theworkingdilutionis1:10.i.e.Add25μlsampleinto225μlsamplediluentbuffer.

Lowtargetproteinconcentration(62.5-4000pg/ml).Theworkingdilutionis1:2.i.e.Add100μlsampleto100μlsamplediluentbuffer.

VeryLowtargetproteinconcentration(≤62.5pg/ml).Nodilutionnecessary,ortheworkingdilutionis1:2.

3.ReagentPreparationandStorage

A.Reconstitutionofthehu

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。