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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但是最好还是不要用,影响实验就不好了。
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
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
我使用的是1%琼脂糖凝胶,电泳缓冲液为0.5×TBE。
请问各位大虾,在不影响实验结果的前提下,这样的含EB的电泳缓冲液可以反复使用多少次?
谢谢^_^
500ml
2。阴极缓冲液 ( 10 × ) :将 60.55g Tris ,89.58g Tricine 及 5g SDS 溶于400ml 蒸馏水中,加水至终体积为500ml
使用时稀释至1×的缓冲液,电泳槽内槽加入阴极电泳缓冲液,外槽加入阳极电泳缓冲液。形成Trcine-SDS-PAGE电泳系统。

