NephrinELISA | |
| Manufacturer/Trade: | Exocell/NephrinELISA |
| CatalogueNumbers: | 1019 |
| Methodology: | CompetitiveELISA |
| SummaryofProcedure: | NephrinELISAisanindirectcompetitiveELISAdesignedtomeasurenephrininurinespecimensofrodentorhumanorigin.Nephrin,atransmembraneproteinexpressedinepithelialcells(podocytes)oftherenalglomerulus,regulatespassageofplasmaproteinsacrosstheglomerularfiltrationbarrierandmaybeshedintotheurineindiseasesaffectingtheglomerulussuchasdiabetesmellitus. Tocompletetheassay,standardorsamplecontainingnephrininthesolublephaseisaddedtomicrotiterwellsthatcontainnephrinantigenimmobilizedontothesolidphase,andtowhichrabbitanti-nephrinantibodyisaddedinthefluidphase.Theantibodybindstoeithertheimmobilizednephrinorthesolublenephrin,whichcompeteforthisbinding.Afteraonehourincubationatroomtemperature,theplatesarewashedandagoatanti-rabbit–HRPconjugateisusedtodetectboundantibody.Platesarewashedandonlytheantibodyconjugateboundtothestationaryphaseantigenremainsinthewell,whichisdetectedwithachromogenicreaction.Colorintensityisinverselyproportionaltotheconcentrationofnephrininthefluidphase.Concentrationsinexperimentalsamplesaredeterminedfromastandardcurve. |
| SpecimenRequired: | Urine,100uL |
| AssayRange: | 0.031-2.0ug/ml |
| Precision: | IntraassayandinterassayprecisionforsamplesintheusefulrangehaveCVs<10% of the mean |
| Specificity: | Rabbitanti-nephrinantibodyreactswithseveralmammalianspeciesincludingrodentandhuman |
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