Description:
CytokinesaresignalingmoleculesthathavecriticalrolesinmanyBIOLOGicalprocessessuchascellulargrowth,differentiation,geneexpression,migration,immunity,andinflammation.Cytokinesthataresecretedfromcellsbindtocell-surfacereceptors, initiatetheactivationofsignaltransductionpathwaysandmediatecelltocellcommunication.Themalfunctionofcytokinesleadstomanydiseases,includingarthritis,acuteandchronicliverdisease,inflammatoryboweldisease,cardiac-relateddiseases,andcancers.Cytokinesarecommonlyworkingtogether ina biologicalordiseaseprocess.Therefore,thecomprehensiveanalysisoftheexpressionofmultiplecytokinesallowseffectiverevealingoftheunderneathmechanismofcytokineaction andthealterationleADIngto diseases.TheHumanCytokineELISAPlateArrayisacolormetricdetectionthatallowsyoutomonitortheabundanceof31humancytokinessimultaneously.Thisfastandsensitiveassay canbeusedforquantitativecomparisonof thesecytokinesamongdifferent samples.ApplicableGrid:
ListofApplicableCytokines
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | TNFa | VEGF | PDGF-BB | IL-10 | TNFa | VEGF | PDGF-BB | IL-10 | TNFa | VEGF | PDGF-BB | IL-10 |
| B | IFNy | EGF | PIGF-1 | FGFb | IFNy | EGF | PLGF-1 | FGFb | IFNy | EGF | PLGF-1 | FGFb |
| C | G-CSF | IL-6 | b-NGF | Leptin | G-CSF | IL-6 | b-NGF | Leptin | G-CSF | IL-6 | b-NGF | Leptin |
| D | GM-CSF | Resistin | SCF | IGF-1 | GM-CSF | Resistin | SCF | IGF-1 | GM-CSF | Resistin | SCF | IGF-1 |
| E | IL-1a | PAI-1 | MCP-1 | TGF-b | IL-1a | PAI-1 | MCP-1 | TGF-b | IL-1a | PAI-1 | MCP-1 | TGF-b |
| F | IL-8 | IL-12 | MIP-1a | Adipo | IL-8 | IL-12 | MIP-1a | Adipo | IL-8 | IL-12 | MIP-1a | Adipo |
| G | IP-10 | IL-13 | IL-2 | IL-17a | IP-10 | IL-13 | IL-2 | IL-17a | IP-10 | IL-13 | IL-2 | IL-17a |
| H | Rantes | Eotaxin-3 | IL-4 | IL-1b | Rantes | Eotaxin-3 | IL-4 | IL-1b | Rantes | Eotaxin-3 | IL-4 | IL-1b |
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.
%20-%20Chelsea2.png)
Data:
AnalysisofCytokineProteinExpressioninTNFa-TreatedandUntreatedHeLawithHumanCytokineELISAPlateArray.HeLacellswerestarvedfor24hourswithserum-freemedium,subsequentlytreatedthecellswithandwithout20ng/ulTNFfor16hours.Theserum-freeconditionedmediawereincubatedontheplatefor1hour.AfterincubatingwithdetectionantibodymixandHRP,theplatewasdetectedwithTMBsubstratebyaplatereader. | |
