![StressMarq/StressXpress® Cortisone CLIA Kit/SKT-206-96/96-well](images/StressMarq/SKT-206_Cortisone_CLIA_Kit_Standard_Curve_Fig2.png)
Overview:
ProductName | CortisoneCLIAKit |
Description | Chemiluminescentmeasurementofcortisone |
SpeciesReactivity | SpeciesIndependent |
Platform | Microplate |
SampleTypes | DriedFecalSamples,Plasma,Saliva,Serum,TissueCultureMedia,Urine |
DetectionMethod | ChemiluminescentAssay |
AssayType | SandwichCLIA(ChemiluminescentImmunoassay) |
Utility | CLIAkitusedtoquantitativelymeasurethecortisonepresentinsamples. |
Sensitivity | 10.6pg/ml |
AssayRange | 78.1-20,000pg/ml |
Precision | IntraAssayPrecision:ThreehumansamplesweredilutedwithAssayBufferandruninreplicatesof20inanassay.ThemeanandprecisionofthecalculatedCortisoneconcentrationswere:Sample1-7902.0pg/mL,7.9%CVSample2-596.1pg/mL,5.7%CVSample3-234.0pg/mL,10.1%CVInterAssayPrecision:ThreehumansamplesweredilutedwithAssayBufferandruninduplicatesinthirteenassaysrunovermultipledaysbythreeoperators.ThemeanandprecisionofthecalculatedCortisoneconcentrationswere:Sample1-7904.0pg/mL,10.0%CVSample2-662.7pg/mL,11.1%CVSample3-262.9pg/mL,12.9%CV |
IncubationTime | 2hours |
NumberofSamples | 37samplesinduplicate |
OtherResources | KitBooklet,MSDS,SteroidSolidExtractionProtocol,SalivaSampleHandlingInstructions |
Properties
StorageTemperature | 4ºC | ||||||||||||||||||||||||||||||||||||
ShippingTemperature | BlueIce | ||||||||||||||||||||||||||||||||||||
ProductType | CLIAKits | ||||||||||||||||||||||||||||||||||||
AssayOverview | TheCortisoneCLIAkitisdesignedtoquantitativelymeasureCortisonepresentinextracteddriedfecalsamples,urine,saliva,andserumsamples.Thiskitmeasurestotalcortisoneinserumandplasmaandinextractedfecalsamples.Acortisonestandardisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.StandardsordilutedsamplesarePipettedintoawhitemicrotiterplatecoatedwithanantibodytocapturerabbitantibodies.Acortisone-peroxidaseconjugateisaddedtothestandardsandsamplesinthewells.Thebindingreactionisinitiatedbytheadditionofapolyclonalantibodytocortisonetoeachwell.Afteratwohourincubationtheplateiswashedandthechemiluminescentsubstrateisadded.Thesubstratereactswiththeboundcortisone-peroxidaseconjugatetoproducelight.ThegeneratedlightisdetectedinamicrotiterplatereadercapableofreADIngluminescence.Theconcentrationofthecortisoneinthesampleiscalculated,aftermakingsuitablecorrectionforthedilutionofthesample,usingsoftwareavailablewithmostplatereaders. | ||||||||||||||||||||||||||||||||||||
KitOverview |
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CiteThisProduct | CortisoneCLIAKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-206) |
BIOLOGicalDescription
AlternativeNames | (8S,9S,10R,13S,14S,17R)-17-Hydroxy-17-(2-hydroxyacetyl)-10,13-dimethyl-1,2,6,7,8,9,12,14,15,16-decahydrocyclopenta[a]phenanthrene-3,11-dioneCLIAKit |
ResearchAreas | CellSignaling |
ScientificBackground | Cortisone(C21H28O5,Kendall’sCompound‘E’)wasidentifiedbyMason,MyersandKendallin1936asCompoundEextractedfrombovinesuprarenalglandtissuethathadthequalitativebutnotquantitativeactivityofcortin.Thepresenceofmultiplecortin-likecompoundsledtheauthorstospeculatethatthestudyofCompoundEwouldrevealthenatureofcortin(1).CompoundEisnowcalledcortisoneandthemoreactiveCompoundF,cortisol,andtheconcentrationsofthesetwoglucocorticoidsvaryduetotheactivityoftwo11ß-hydroxysteroiddehydrogenases(11-HSD)(2,3).WhilemosttissueshavetheABIlitytoexpresseitherenzyme,11ß-HSD1isfoundprimarilyintheliverwhereitconvertscortisonetocortisolwhile11ß-HSD2isfoundintissuessuchasthekidneywherecortisolreceptorbindingisrequired.11ß-HSD2deactivatescortisoltocortisone,prohibitingreceptoractivation.Thisglucocorticoid“shuttle”helpstoinitiateandregulatetheanti-inflammatoryresponse,makingcortisoneoneofthemodern“wonderdrugs”.onitoringtheratioofcortisone:cortisolhasapplicationsindiabetes,obesity,metabolicsyndrome,osteoporosis,andchronicfatiguesyndromeinadditiontoadrenaldiseases(4-7).Cortisoneandcortisolconcentrationsexhibitapredictablediurnalpatternandcanbemeasuredinextracteddriedfeces,orinserum,plasma,salivaandurine.Arecentpublication(8)hassuggestedthatsalivarycortisoneisagoodsurrogateMarkerforserumcortisol. |
References | 1.Mason,HL,etal.J.Biol.Chem.,1936116:267-276. 2.Mason,HL,et.al.J.Biol.Chem.,1938124:459-474. 3.Hillier,SG.“DiamondsareForever:theCortisoneLegacy”J.Endo.,2007195:1-6. 4.vanRaalte,DH,etal.Eur.J.Clin.Invest.200939(2):81-93. 5.Pierotti,S,etal.J.SteroidBiochem.Mol.Biol.2008108(3-5):292-9. 6.Hadoke,PWF,etal.Br.J.Pharmacol.2009156:689-712. 7.Jerkes,WK,etal.J.PsychosomaticRes.200660:145-153. 8.Perogamvros,I,etal.JClin.Endocrin.Metab.2010August4(Epubaheadofprint). |
ProductImages
![<p>Typical Standard Curve for the Cortisone CLIA Kit (Chemiluminescent Immunoassay) StressXpress® – SKT-206. Assay Type: Sandwich CLIA. Detection Method: Chemiluminescent Assay. Assay Range: 78.1 – 20,000 pg/ml.</p>](images/StressMarq/201710/SKT-206_Cortisone_CLIA_Kit_Standard_Curve_Fig2.png)
TypicalStandardCurvefortheCortisoneCLIAKit(ChemiluminescentImmunoassay)StressXpress®–SKT-206.AssayType:SandwichCLIA.DetectionMethod:ChemiluminescentAssay.AssayRange:78.1–20,000pg/ml.
![<p>Linearity was determined by taking two serum samples treated with Dissociation Reagent and diluted 1:50 with Assay Buffer, one with a low diluted cortisone level of 1,336 pg/mL and one with a higher diluted level of 4,055 pg/mL, and mixing them. The measured concentrations were compared to the expected values based on the ratios used.</p>](images/StressMarq/201710/SKT-206_Cortisone_CLIA_Kit_Linearity_Recovery_Graph_Fig3.png)
LinearitywasdeterminedbytakingtwoserumsamplestreatedwithDissociationReagentanddiluted1:50withAssayBuffer,onewithalowdilutedcortisonelevelof1,336pg/mLandonewithahigherdilutedlevelof4,055pg/mL,andmixingthem.Themeasuredconcentrationswerecomparedtotheexpectedvaluesbasedontheratiosused.
![<p>Chemical structure of Cortisone</p>](images/StressMarq/201710/SKT-206_Cortisone_CLIA_Kit_Chemical_Structure_of_Cortisone_Fig1.png)
ChemicalstructureofCortisone
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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
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。
Tris-Glycine/SDS;
MOPS/SDS;
Bis-Glycine/SDS等不同的缓冲液、预混液等。
不知道这几种缓冲液有什么异同呢?
比如如果是Invitrogen的预制胶-预混液电泳系统,每种预制胶有适配缓冲液,但是如果用其他的缓冲液似乎也可以得到不错的结果,那么是否说明电泳缓冲液只要满足了缓冲ph范围,其他的不是特别重要呢?
此外,附加提问是,PVDF膜在转膜的时候,转膜缓冲液中不需要加入甲醇,那么是加入甲醇转膜效果好还是不加好呢?
谢谢!
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