
Overview:
ProductName | UrineCreatinineDetectionKit |
Description | Quantitativecolorimetricdetectionofcreatinineinurinesamples |
SpeciesReactivity | Dog,Human,Monkey,Rat |
Platform | Microplate |
SampleTypes | Urine |
DetectionMethod | ColorimetricAssay |
AssayType | DirectQuantitativeAssay |
Utility | Detectionkitusedtoquantitativelymeasurecreatinineinsamples. |
Sensitivity | 0.019mg/dl |
AssayRange | 0.3125-20mg/dl |
Precision | IntraAssayPrecision:Threehumanurinesampleswerediluted1:20withdeionizedwaterandruninreplicatesof20inanassay.Themeanandprecisionofthecalculatedcreatinineconcentrationswere:Sample1-1174.3pg/mL,6%CVSample2-475.9pg/mL,5.6%CVSample3-177.4pg/mL,14.7%CVInterAssayPrecision:Threehumanurinesampleswerediluted1:20withdeionizedwaterandruninduplicatesin20assaysrunoverfivedaysbythreeoperators.Themeanandprecisionofthecalculatedcreatinineconcentrationswere:Sample1-1188.1pg/mL,7.2%CVSample2-508.7pg/mL,6.3%CVSample3-199.7pg/mL,10.9%CV |
NumberofSamples | 88samplesinduplicate |
OtherResources | KitBooklet,MSDS |
Properties
StorageTemperature | 4ºC | |||||||||||||||
ShippingTemperature | BlueIce | |||||||||||||||
ProductType | DetectionKits | |||||||||||||||
AssayOverview | TheUrineCreatinineDetectionkitisdesignedtoquantitativelymeasurecreatininepresentinurinesamples.Acreatininestandard,calibratedtoaNISTcreatininestandard,isprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.StandardsordilutedsamplesarePipettedintoaclearmicrotiterplate.ThecolorgeneratingreactionisinitiatedwiththeStressXpress®CreatinineReagent,whichispipettedintoeachwell.Afterashortincubationtheintensityofthegeneratedcolorisdetectedinamicrotiterplatereadercapableofmeasuring490nmwavelength.Theconcentrationofthecreatineinthesampleiscalculated,aftermakingasuitablecorrectionforthedilutionofthesample,usingsoftwareavailablewithmostplatereaders.TheJaffereactionusedinthiskithasbeenmodifiedtoreadcreatininelevelsinurine(7,8). | |||||||||||||||
KitOverview |
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CiteThisProduct | UrineCreatinineDetectionKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-200) |
BIOLOGicalDescription
AlternativeNames | N-Carbamimidoyl-N-methylglycineDetectionKit,MethylguanidoaceticacidDetectionKit |
ResearchAreas | CardiovascularSystem,CellSignaling |
ScientificBackground | Creatinine(2-amino-1-methyl-5H-imadazol-4-one)isametaboliteofphosphocreatine(p-creatine),amoleculeusedasastoreforhigh-energyphosphatethatcanbeutilizedbytissuesfortheproductionofATP(1).Creatineeithercomesfromthedietorsynthesizedfromtheaminoacidsarginine,glycine,andmethionine.Thisoccursinthekidneysandliver,althoughotherorgansystemsmaybeinvolvedandspecies-specificdifferencesmayexist(2).Creatineandp-creatineareconvertednon-enzymaticallytothemetabolitecreatinine,whichdiffusesintothebloodandisexcretedbythekidneys.Invivo,thisconversionappearstobeirreversIBLeandinvitroitisfavoredbyhighertemperaturesandlowerpH2.Creatinineformsspontaneouslyfromp-creatine(3).Undernormalconditions,itsformationoccursataratethatisrelativelyconstantandasintra-individualvariationis<15% from day to day, creatinine is a useful tool for normalizing the levels of other molecules found in urine. Additionally altered creatinine levels may be associated with other conditions that result in decreased renal blood flow such as diabetes and cardiovascular disease (4-6). |
References | 1.Wallimann,T.etal.,Biochem.J.,2000,281,21-40. 2.Wyss,M.andKaddurah-Daouk,R.,Physiol.Rev.,2000,80,1107-1213. 3.RajaIyengar,M.etal.,J.Biol.Chem,1985,260,7562-7567. 4.Manjunath,G.etal.,Postgrad.Med.2001,110,55-62. 5.Gross,J.L.etal.,DiabetesCare,2005,28,164-176. 6.Anavekar,N.S.etal.,NewEngl.J.Med.,2004,351,1285-1295. |
ProductImages

TypicalStandardCurveofUrineCreatinineDetectionKitStressXpress®–SKT-200.AssayType:Quantitative.DetectionMethod:ColorimetricAssay.AssayRange:0.3125-20mg/dl

Conversionofcreatineandp-creatinetocreatinine.Creatineandp-creatineareconvertednon-enzymaticallytothemetabolitecreatinine,whichdiffusesintothebloodandisexcretedbythekidneys.Invivo,thisconversionappearstobeirreversibleandinvitroitisfavoredbyhighertemperaturesandlowerpH.Creatinineformsspontaneouslyfromp-creatine.

Linearitywasdeterminedbytakingtwodiluted1:20humanurinesamples,onewithalowdilutedcreatininelevelof0.38mg/dLandonewithahigherdilutedlevelof9.33mg/dLandmixingthemintheratiosgivenbelow.Themeasuredconcentrationswerecomparedtotheexpectedvalues.
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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。

