
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。
低离子强度时,迁移率快。但离子强度过低,缓冲液的缓冲容量小,不易维持pH恒定。高离子强度时迁移率慢,因此为了获得更快的反应速度,在缓冲容量允许的范围内,离子强度应该尽可能小。
2.配好的EDTA不灭菌行不?放4度冰箱等过完年回来直接拿去配TAE
3.TAE配好后不用灭菌吧
TAE是使用最广泛的缓冲系统。其特点是超螺旋在其中电泳时更符合实际相对分子质量(TBE中电泳时测出的相对分子质量会大于实际分子质量),且双链线状DNA在其中的迁移率较其他两种缓冲液快约10%,电泳大于13kb的片段时用TAE缓冲液将取得更好的分离效果,此外,回收DNA片段时也易用TAE缓冲系统进行电泳。TAE的缺点是缓冲容量小,长时间电泳(如过夜)不可选用,除非有循环装置使两极的缓冲液得到交换。 50×TAE Buffer 配制方法: 1. 称量Tris 242g,Na2EDTA.2H2O 37.2g 于1L烧杯中; 2.向烧杯中加入约800ml去离子水,充分搅拌均匀; 3加入57.1ml的冰乙酸,充分溶解; 4.加去离子水定容至1L后,室温保存。向左转|向右转
但是最好还是不要用,影响实验就不好了。

