
Description:
Mouseleptinplaysakeyroleinregulatingenergyintakeandenergyexpenditure,includingtheregulation(decrease)ofappetiteand(increase)ofmetabolism. LeptinisproducedbyADIposetissueandthelevelofcirculatingleptinisdirectlyproportionaltothetotalamountoffatinthebody.OnceleptinhasboundtotheOb-Rbreceptor,itactivatesStat3,whichisphosphorylatedandtravelstothenucleuswhereitmediatesgeneexpression.Oneofthemaineffectsongeneexpressionisthedown-regulationoftheexpressionofendocannABInoids,responsIBLe—amongtheirmanyotherfunctions—forincreasingappetite.Thereareotherintracellularpathwaysactivatedbyleptin,butlessisknownabouthowtheyfunctioninthissystem.Althoughleptinisacirculatingsignalthatreducesappetite,ingeneral,obesepeoplewhoareresistanttotheeffectsofleptinhaveanunusuallyhighcirculatingconcentrationofleptin,inthesamewaythatpeoplewithtype2diabetesareresistanttotheeffectsofinsulin. Leptinisalsofoundtostimulateendothelialcellproliferationandangiogenesis.Principle:
Mouse CytokineELISAisbasedontheprincipleofasolidphaseenzyme-linkedimmunosorbentassay.Theassayutilizesrabbitanti-mouseantibodiesforimmobilizationonthemicrotiterwellsandrabbitanti-mouseantibodiesalongwithstreptavidinconjugatedtohorseradishperoxidase(HRP)fordetection.Thetestsampleisallowedtoreactsimultaneouslywiththetwoantibodies,resultinginthemoleculesbeingsandwichedbetweenthesolidphaseandenzyme-linkedantibodies.Afterincubation,thewellsarewashedtoremoveunbound-labeledantibodies.AHRPsubstrate,TMB,isaddedtoresultinthedevelopmentofabluecolor.ThecolordevelopmentisthenstoppedwiththeadditionofStopSolutionchangingthecolortoyellow.The concentrationofVEGFisdirectlyproportionaltothecolorintensityofthetestsample.Absorbanceismeasuredspectrophotometricallyat450nm.
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