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 |
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) |
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. |
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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