Overview:
| ProductName | CorticosteroneEIAKit |
| Description | Colorimetricdetectionofcorticosterone |
| SpeciesReactivity | SpeciesIndependent |
| Platform | Microplate |
| SampleTypes | DriedFecalSamples,EDTAPlasma,HeparinPlasma,Serum,TissueCultureMedia,Urine |
| DetectionMethod | ColorimetricAssay |
| AssayType | SandwichEIA(EnzymeImmunoassay) |
| Utility | EIAkitusedtomeasurethecorticosteronepresentinsamples. |
| Sensitivity | 18.6pg/ml |
| AssayRange | 78.125-10,000pg/ml |
| Precision | IntraAssayPrecision:FourhumansamplesweredilutedwithAssayBufferandruninreplicatesof20inanassay.ThemeanandprecisionofthecalculatedCorticosteroneconcentrationswere:Sample1-2460.6pg/mL,6.3%CVSample2-601.5pg/mL,6.5%CVSample3-371.6pg/mL,3.1%CVSample4-259.0pg/mL,4.8%CVInterAssayPrecision:ThreehumansamplesweredilutedwithAssayBufferandruninduplicatesinfourteenassaysrunovermultipledaysbyfouroperators.ThemeanandprecisionofthecalculatedCorticosteroneconcentrationswere:Sample1-2618.3pg/mL,7.5%CVSample2-630.1pg/mL,6.4%CVSample3-267.9pg/mL,9.9%CV |
| NumberofSamples | 39samplesinduplicate |
| OtherResources | KitBooklet,MSDS,SteroidSolidExtractionProtocol |
Properties
| StorageTemperature | 4ºC | |||||||||||||||||||||||||||||||||
| ShippingTemperature | BlueIce | |||||||||||||||||||||||||||||||||
| ProductType | EIAKits | |||||||||||||||||||||||||||||||||
| AssayOverview | TheCorticosteroneEIAkitisdesignedtoquantitativelymeasureCorticosteronepresentinserum,plasma,urine,extracteddriedfecalsamples,andtissueculturemediasamples.Thiskitmeasurestotalcorticosteroneinserumandplasmaandinextractedfecalsamples.Acorticosteronestocksolutionisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.Weprovideprotocolsonpage8toprepareassaystandardsfrom5,000to78.125pg/mLorfrom10,000to78.125pg/mL.Pleasechoosethestandardrangethatfitsyoursampleconcentrationsmostappropriately.StandardsordilutedsamplesarePipettedintoaclearmicrotiterplatecoatedwithanantibodytocapturesheepantibodies.Acorticosterone-peroxidaseconjugateisaddedtothestandardsandsamplesinthewells.Thebindingreactionisinitiatedbytheadditionofapolyclonalantibodytocorticosteronetoeachwell.Afteranhourincubationtheplateiswashedandsubstrateisadded.Thesubstratereactswiththeboundcorticosterone-peroxidaseconjugate.Afterashortincubation,thereactionisstoppedandtheintensityofthegeneratedcolorisdetectedinamicrotiterplatereadercapableofmeasuring450nmwavelength.Theconcentrationofthecorticosteroneinthesampleiscalculated,aftermakingsuitablecorrectionforthedilutionofthesample,usingsoftwareavailablewithmostplatereaders. | |||||||||||||||||||||||||||||||||
| KitOverview |
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| CiteThisProduct | CorticosteroneEIAKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-205) |
BIOLOGicalDescription
| AlternativeNames | (11β)-11,21-dihydroxypregn-4-ene-3,20-dioneEIAKit |
| ResearchAreas | Cancer,CellSignaling,Neuroscience,OxidativeStress |
| ScientificBackground | Corticosterone(C21H30O4,Kendall’sCompound‘B’)isaglucocorticoidsecretedbythecortexoftheadrenalgland.CorticosteroneisproducedinresponsetostimulationoftheadrenalcortexbyACTHandistheprecursorofaldosterone.Corticosteroneisamajorindicatorofstressandisthemajorstresssteroidproducedinnon-humanmammals.Studiesinvolvingcorticosteroneandlevelsofstressincludeimpairmentoflongtermmemoryretrieval(1),chroniccorticosteroneelevationduetodietaryrestrictions(2)andinresponsetoburninjuries(3).Inadditiontostresslevels,corticosteroneisbelievedtoplayadecisiveroleinsleep-wakepatterns(4,5). |
| References | 1.Hupé,JM,etal.Nature,1998;394:784-787. 2.KitayskyAS,KitaiskaiaEV,WingfieldJC,PiattJF.J.Comp.Physiol,2001;171:701-709. 3.ThellinO,NoelG,KhuanaS,OgleCKandHorsemanND.Shock,2001;16(5):393-397. 4.Krame,KM.andSothernRB.Chronobiol.Int.,2001;18(6):933-945. 5.Vazquez-PalaciosG,etal.Pharmacol.BiochemBehavior,2001:70(2-3):305-310. |
ProductImages

TypicalStandardCurvefortheCorticosteroneEIAKit(EnzymeImmunoassay)StressXpress®–SKT-205.AssayType:SandwichEIA.DetectionMethod:ColorimetricAssay.AssayRange:78.125–10,000pg/ml.

LinearitywasdeterminedbytakingtwoserumsamplestreatedwithDissociationReagentanddiluted1:50withAssayBuffer,onewithalowdilutedcorticosteronelevelof104.6pg/mLandonewithahigherdilutedlevelof2,890.5pg/mL,andmixingthemintheratiosgivenbelow.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.

ChemicalstructureofCorticosterone
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② 应取pH8.0,这样可使核苷酸带较多负电荷,利于吸附于阴离子交换树脂柱。虽然pH 11.4时核苷酸带有更多的负电荷,但pH过高对分离不利。
③ 当不考虑树脂的非极性吸附时,根据核苷酸负电荷的多少来决定洗脱速度,则洗脱顺序为CMP>AMP> GMP > UMP,但实际上核苷酸和聚苯乙烯阴离子交换树脂之间存在着非极性吸附,嘌呤碱基的非极性吸附是嘧啶碱基的3倍。静电吸附与非极性吸附共同作用的结果使洗脱顺序为:CMP> AMP > UMP >GMP。
1. Buffer中离子浓度过大,甘氨酸或者Tris碱有可能疏忽多加了
2.没有加相应浓度的SDS
3.电泳周围温度高,也是一个原因

