Description:
CytokinesaresignalingmoleculesthathavecriticalrolesinmanyBIOLOGicalprocessessuchascellulargrowth,differentiation,geneexpression,migration,immunity,andinflammation.Cytokinesaresecretedfromcellsandboundtocell-surfacereceptors,whichinitiatetheactivationofsignaltransductionpathwaysandmediatecelltocellcommunication.Themalfunctionofcytokinesleadstomanydiseases,includingarthritis,acuteandchronicliverdisease,inflammatoryboweldisease,cardiac-relateddiseases,andcancers.Agroupofcytokinescommonlyinvolvesinonebiologicalordiseaseprocess.Therefore,thecomprehensiveanalysisoftheexpressionofmultiplecytokinesallowsrevealingtheunderneathmechanismofthediseasestateeffectively.TheRatCytokineELISAPlateArrayallowsyoutomonitortheabundanceof16ratcytokinesinahigh-throughputmanner.Thisassayisafastandsensitivetoolforquantitativelyprofilingthelevelsofmultiplecytokinesbetweensamplessimultaneously.Principle:
The96-wellwhiteplateisdividedinto3or4sections,andeachsectionhas4or3columnsforonesample.Ineachsection,32(HumanCytokineELISAPlateArrays)or24(MouseCytokineELISAPlateArray)specificcytokinecaptureantibodiesarecoatedon32or24wellsrespectively.Thesample,cellculturesupernatants,celllysates,tissuehomogenates,serum,orplasmasamplesamongothers areincubatedwiththecytokineELISAplate,and thecapturedcytokineproteinsaresubsequentlydetectedwithacocktailofbiotinylateddetectionantibodies.Thetestsampleisallowedtoreactwith apairof antibodies,resultinginthecytokinesbeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.TheplateisfurtherdetectedwithHRPluminescentsubstrateorHRPsubstrateTMB.Thelevelofexpressionforeachspecificcytokineisdirectlyproportionaltotheluminescentorcolor intensity.
Data:
AnalysisofCytokineProteinExpressioninTNFa-TreatedandUntreatedwithRatCytokineELISAPlateArray.CWSV-1Ratlivercellswerestarvedfor24hourswithserum-freemedium,subsequentlytreatedthecellswithandwithout20ng/ulTNFfor16hours.Theserum-freeconditionedmediawereincubatedontheplatefor1hour.AfterincubatingwithdetectionantibodymixandHRP,theplatewasdetectedwithchemilumincentsubstratebyaplatereader. | |