Description:
CytokinesaresignalingmoleculesthathavecriticalrolesinmanyBIOLOGicalprocessessuchascellulargrowth,differentiation,geneexpression,migration,immunity,andinflammation.Cytokinesthataresecretedfromcellsbindtocell-surfacereceptors, initiatetheactivationofsignaltransductionpathwaysandmediatecelltocellcommunication.Themalfunctionofcytokinesleadstomanydiseases,includingarthritis,acuteandchronicliverdisease,inflammatoryboweldisease,cardiac-relateddiseases,andcancers.Cytokinesarecommonlyworkingtogether ina biologicalordiseaseprocess.Therefore,thecomprehensiveanalysisoftheexpressionofmultiplecytokinesallowseffectivelyrevealingtheunderneathmechanismofcytokineaction andthealterationleADIngto diseases.TheMouseCytokineELISAPlateArray(Colorimetric)allowsyoutomonitortheabundanceof24mousecytokinessimultaneously.Thisfastandsensitiveassay canbeusedforquantitativecomparisonof thesecytokinesamongdifferent samples.ApplicableGrid:
ListofApplicableCytokines
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | TNFa | IL-1a | PDGF-BB | TNFa | IL-1a | PDGF-BB | TNFa | IL-1a | PDGF-BB | TNFa | IL-1a | PDGF-BB |
| B | IGF-1 | IL-1b | b-NGF | IGF-1 | IL-1b | b-NGF | IGF-1 | IL-1b | b-NGF | IGF-1 | IL-1b | b-NGF |
| C | VEGF | G-CSF | IL-17A | VEGF | G-CSF | IL-17A | VEGF | G-CSF | IL-17A | VEGF | G-CSF | IL-17A |
| D | IL-6 | GM-CSF | IL-2 | IL-6 | GM-CSF | IL-2 | IL-6 | GM-CSF | IL-2 | IL-6 | GM-CSF | IL-2 |
| E | FGFb | MCP-1 | IL-4 | FGFb | MCP-1 | IL-4 | FGFb | MCP-1 | IL-4 | FGFb | MCP-1 | IL-4 |
| F | IFNy | MIP-1a | IL-10 | IFNy | MIP-1a | IL-10 | IFNy | MIP-1a | IL-10 | IFNy | MIP-1a | IL-10 |
| G | EGF | SCF | Resistin | EGF | SCF | Resistin | EGF | SCF | Resistin | EGF | SCF | Resistin |
| H | Leptin | Rantes | IL-12 | Leptin | Rantes | IL-12 | Leptin | Rantes | IL-12 | Leptin | Rantes | IL-12 |
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.
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Data:
AnalysisofCytokineProteinExpressioninTNFa-TreatedandUntreatedNIH3T3withMouseCytokineELISAPlateArray.NIH/3T3cellswerestarvedfor24hourswithserum-freemedium,subsequentlytreatedthecellswithandwithout20ng/ulTNFfor16hours.Theserum-freeconditionedmediawereincubatedontheplatefor1hour.AfterincubatingwithdetectionantibodymixandHRP,theplatewasdetectedwithchemilumincentsubstratebyaplatereader. |
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