
Overview:
ProductName | CortisolEIAKit |
Description | Quantitativecolorimetricdetectionofcortisol |
SpeciesReactivity | SpeciesIndependent |
Platform | Microplate |
SampleTypes | DriedFecalSamples,EDTAPlasma,HeparinPlasma,Saliva,Serum,TissueCultureMedia,Urine |
DetectionMethod | ColorimetricAssay |
AssayType | SandwichEIA(EnzymeImmunoassay) |
Utility | EIAkitusedtoquantitativelymeasurecortisolpresentinsamples. |
Sensitivity | 17.3pg/ml |
AssayRange | 100-3200pg/ml |
Precision | IntraAssayPrecision:ThreehumansamplesweredilutedwithAssayBufferandruninreplicatesof20inanassay.ThemeanandprecisionofthecalculatedCortisolconcentrationswere:Sample1-1174.3pg/mL,6%CVSample2-475.9pg/mL,5.6%CVSample3-177.4pg/mL,14.7%CVInterAssayPrecision:ThreehumansamplesweredilutedwithAssayBufferandruninduplicatesintenassaysrunovermultipledaysbyfouroperators.ThemeanandprecisionofthecalculatedCortisolconcentrationswere:Sample1-1188.1pg/mL,7.2%CVSample2-508.7pg/mL,6.3%CVSample3-199.7pg/mL,10.9%CV |
IncubationTime | 90Minutes |
NumberofSamples | 40samplesinduplicate |
OtherResources | KitBooklet,MSDS,SteroidSolidExtractionProtocol,SalivaSampleHandlingInstructions |
Properties
StorageTemperature | 4ºC | |||||||||||||||||||||||||||||||||
ShippingTemperature | BlueIce | |||||||||||||||||||||||||||||||||
ProductType | EIAKits | |||||||||||||||||||||||||||||||||
AssayOverview | TheCortisolEIAkitisdesignedtoquantitativelymeasurecortisolpresentindriedfecalextracts,saliva,urine,serum,plasmaandtissueculturemediasamples.Thiskitmeasurestotalcortisolinextractedsamplesandinserumandplasmaandfreecortisolinsalivaandurine.Acortisolstandardisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.StandardsordilutedsamplesarePipettedintoaclearmicrotiterplatecoatedwithanantibodytocapturemouseantibodies.Acortisol-peroxidaseconjugateisaddedtothestandardsandsamplesinthewells.Thebindingreactionisinitiatedbytheadditionofamonoclonalantibodytocortisoltoeachwell.Afteran1hourincubationtheplateiswashedandsubstrateisadded.Thesubstratereactswiththeboundcortisol-peroxidaseconjugate.Afterashortincubation,thereactionisstoppedandtheintensityofthegeneratedcolorisdetectedinamicrotiterplatereadercapableofmeasuring450nmwavelength.Theconcentrationofthecortisolinthesampleiscalculated,aftermakingsuitablecorrectionforthedilutionofthesample,usingsoftwareavailablewithmostplatereaders. | |||||||||||||||||||||||||||||||||
KitOverview |
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CiteThisProduct | CortisolEIAKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-201) |
BIOLOGicalDescription
AlternativeNames | (11β)-11,17,21-trihydroxypregn-4-ene-3,20-dioneEIAKit |
ResearchAreas | Cancer,CellSignaling,Neuroscience,OxidativeStress |
ScientificBackground | Cortisol,C21H30O5,(hydrocortisone,compoundF)istheprimaryglucocorticoidproducedandsecretedbytheadrenalcortex.Itisoftenreferredtoasthe“stresshormone”asitisinvolvedintheresponsetostressanditaffectsbloodpressure,bloodsugarlevels,andotheractionsofstressadaptation.Immunologically,cortisolfunctionsasanimportantanti-inflammatoryandplaysaroleinhypersensitivity,immunosuppression,anddiseaseresistance(1).Inthemetabolicaspect,cortisolpromotesgluconeogenesis,liverglycogendeposition,andthereductionofglucoseutilization(2).ProductionofcortisolfollowsanACTH-dependentcircADIanrhythm,withapeaklevelinthemorninganddecreasinglevelsthroughouttheday.Mostserumcortisol,allbutabout4%,isboundtoproteinsincludingcorticosteroidbindingglobulinandserumalbumin(1,3).Onlyfreecortisolisavailabletomostreceptorsanditisthroughthesereceptorsthatphysiologicalprocessesaremodulated.Abnormalcortisollevelsarebeingevaluatedforcorrelationwithavarietyofdifferentconditions,suchasprostatecancer(4),depression(5),andschizophrenia(6).ItisalreadyknownthatabnormallevelsofcortisolareinvolvedinCushing’sSyndromeandAddision’sdisease(7). |
References | 1.E.Friess,etal.,EurJClinInvest,2000,30,Suppl3:46-50. 2.Freeman,Scott,2002.BiologicalScience.PrenticeHall;2ndPkgedition(December30,2004). 3.C.Longscope.,J.Endocrinology,1996,,SupplS125-S127. 4.J.Herbert,Lancet,1995345,1193-1194. 5.A.Michael,etal.,Biol.Psychiatry,2000,48,989-95. 6.C.R.DequetandD.J.Wallace,CurrentOpin.Ivest.Drugs,2001,8,1045-53. 7.W.M.Jeffries,Med.Hypotheses,1998,51,114-4. |
ProductImages

TypicalStandardCurvefortheCortisolEIAKit(EnzymeImmunoassay)StressXpress®–SKT-201.AssayType:SandwichEIA.DetectionMethod:ColorimetricAssay.AssayRange:100–3200pg/ml.

Linearitywasdeterminedbytakingtwohumanurinesamplesdiluted1:140,onewithalowdilutedcortisollevelof163.9pg/mLandonewithahigherdilutedlevelof2,974.9pg/mLandmixingthemintheratiosgivenbelow.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.
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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。

