
Overview:
ProductName | CystatinCEIAKit |
Description | ColorimetricmeasurementofhumancystatinClevels |
SpeciesReactivity | Human |
Platform | Microplate |
SampleTypes | EDTAPlasma,HeparinPlasma,Serum,TissueCultureMedia,Urine |
DetectionMethod | ColorimetricAssay |
AssayType | SandwichEIA(EnzymeImmunoassay) |
Utility | EIAkitusedtomeasurecystatinClevelsinsamples. |
Sensitivity | 0.058ng/ml |
AssayRange | 0.156-10ng/ml |
Precision | IntraAssayPrecision:FourhumansamplesweredilutedwithAssayBufferandruninreplicatesof20inanassay.ThemeanandprecisionofthecalculatedCystatinCconcentrationswere:Sample1-7.72ng/mL,9.1%CVSample2-5.08ng/mL,10.3%CVSample3-4.42ng/mL,7.6%CVSample4-0.88ng/mL,10.3%CVInterAssayPrecision:FourhumansamplesweredilutedwithAssayBufferandruninduplicatesintenassaysrunovermultipledaysbythreeoperators.ThemeanandprecisionofthecalculatedCystatinCconcentrationswere:Sample1-7.79ng/mL,8.4%CVSample2-5.48ng/mL,10.2%CVSample3-4.97ng/mL,11.1%CVSample4-1.04ng/mL,12.40%CV |
IncubationTime | 2hours |
NumberofSamples | 40samplesinduplicate |
OtherResources | KitBooklet,MSDS |
Properties
StorageTemperature | 4ºC | |||||||||||||||||||||||||||
ShippingTemperature | BlueIce | |||||||||||||||||||||||||||
ProductType | EIAKits | |||||||||||||||||||||||||||
AssayOverview | TheCystatinCEIAkitisdesignedtoquantitativelymeasurehumanCystatinCpresentinBIOLOGicalsamplesandtissueculturemedia.AhumanCystatinCstandardisprovidedtogenerateastandardcurvefortheassayandallsamplesshouldbereadoffthestandardcurve.StandardsordilutedsamplesarePipettedintoaclearmicrotiterplatecoatedwithamonoclonalantibodytocapturetheCystatinCpresent.Aftera60minuteincubation,theplateiswashedandaperoxidaseconjugatedCystatinCmonoclonalantibodyisadded.Theplateisagainincubatedfor30minutesandwashed.Substrateisthenaddedtotheplate,whichreactswiththeboundCystatinCAntibodyConjugate.Afterathirdincubation,thereactionisstoppedandtheintensityofthegeneratedcolorisdetectedinamicrotiterplatereadercapableofmeasuring450nmwavelength.TheconcentrationoftheCystatinCinthesampleiscalculated,aftermakingsuitablecorrectionfordilution,usingsoftwareavailablewithmostplatereaders. | |||||||||||||||||||||||||||
KitOverview |
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CiteThisProduct | CystatinCEIAKit(StressMarqBiosciencesInc.,VictoriaBCCANADA,Catalog#SKT-219) |
BiologicalDescription
AlternativeNames | Cystatin3EIAKit,gammatraceEIAKit,post-gamma-globulinorneuroendocrinebasicpolypeptideEIAKit |
ResearchAreas | Cancer,Alzheimer"sDisease,CellSignaling,Neurodegeneration,Neuroscience |
ScientificBackground | CystatinCisanon-glycosylatedproteinoflowmolecularweight(13kDa)inthecystatinsuperfamily.CystatinCisproducedataconstantrateinallnucleatedcells,secretedfromcellsandthusfoundindetectableamountsinmostbodyfluids(1,2).CystatinCbelongstothecysteineproteinaseinhibitorgroupandisassociatedwithseveralpathologicalconditions.ImbalancebetweenCystatinCandcysteineproteinasesisassociatedwithdiseasessuchasinflammation,renalfailure,cancer,Alzheimer’sdisease,multiplesclerosisandhereditaryCystatinCamyloidangiopathy(1,3,4).Increasedlevelshavebeenfoundinautoimmunediseases,withcolorectaltumorsandmetastases,patientswithinflammationandinpatientsondialysis(5,6,7).CystatinCisremovedfrombloodplasmabyglomerularfiltrationinthekidneys.Itisreabsorbedbytheproximaltubularcellsanddegraded.ThereisalinearrelationshipbetweenthereciprocalCystatinCconcentrationinplasmaandtheglomerularfiltrationrate(GFR).CystatinCissuggestedtobeabetterMarkerforGFRthantheubiquitousserumcreatininemarkerasitsserumconcentrationisnotaffectedbyotherfactorssuchasage,genderandbodymassandCystatinChashighersensitivitytodetectareducedGFRthancreatininedetermination(2,5).LowlevelsofCystatinCarefoundwiththebreakdownoftheelasticlaminaeandatherosclerosisandaBDominalaorticaneurysm(8).ThereisevidentassociationofCystatinClevelswiththeincidenceofmyocardialinfarction,coronarydeathandanginapectoris,presentingariskfactorforsecondarycardiovascularevents(9). |
References | 1.PergandeM.andJungK.ClinChem.199339(9):1885-90. 2.NewmanD,etal.KidneyInt’l.1995Jan;47(1):312-8. 3.LevyE.ExpertRev.Neurotherapeutics2008May;8(5):687-9. 4.NakashimaI,etal.AnnNeurol.2007Aug;62(2):197-200. 5.UchidaK.andGotoh,A.ClinChimActa.2002Sep;323:121-8. 6.PucciL,etal.ClinChem.2007Mar;53(3):480-8. 7.StabucB,etal.ClinChem.2000Feb;46(2):193-7. 8.ShlipakM,etal.NEJM2005May;352(20):2049-60. 9.KoenigW,etal.ClinChem.2005Feb;51(2):321-7. |
ProductImages

TypicalStandardCurvefortheCystatinCEIAKit(EnzymeImmunoassay)StressXpress®–SKT-219.AssayType:SandwichEIA.DetectionMethod:ColorimetricAssay.AssayRange:0.156–10ng/ml.

Serumlinearitywasdeterminedbytakingtwohumanserumsamplesdiluted1:100,onewithalowdilutedCystatinClevelof4.94ng/mLandonewithahigherdilutedlevelof12.31ng/mLandmixingthemintheratiosgivenbelow.Urinelinearitysampleswerediluted1:16.Onesamplewithavalueof1.64ng/mLwasmixedinratioswithasampleof3.72ng/mL.Thedilutedserumandurinesamplesweremixed4:1,3:2,2:3and1:4andrunintheassay.Themeasuredconcentrationswerecomparedtothevaluespreviouslydetermined.
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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。

