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GenWay/17 beta-ESTRADIOL/GWB-A07AF3/1x96 Assays

Description:

ProductName:17β-EstrADIol

#ofSamples:1x96Assays

IntendedUse:Competitiveimmunoenzymaticcolorimetricmethodforquantitativedeterminationof17β-Estradiolinhumanserumorplasma.

Introduction:Estradiol(17β-Estradiol)isasexhormone.ItrepresentsthemajorEstrogeninhumans.Estradiolhasnotonlyacriticalimpactonreproductiveandsexualfunctioning,butalsoaffectsotherorgansincludingbonestructure.
DuringthereproductiveyearsmostEstradiolinwomenisproducedbytheovaries,smalleramountsofEstradiolarealsoproducedbytheadrenalcortex.Inmen,thetestesproduceEstradiol.
InplasmaEstradiolislargelyboundtosexhormonebindingglobulin(SHBG),alsotoalbumin,onlyafractionisfreeandBIOLOGicallyactive.
SerumEstradiolmeasurementinwomenreflectsprimarilytheactivityoftheovaries.DuringpregnancyEstrogenlevels,includingEstradiol,risesteadilytowardsterm.Estradiolincreasesduetoplacentalproduction.
Inadultpremenopausalwomen,ovarianproductionofEstradiolisstimulatedbyluteinizinghormone(LH)andthefollicle-stimulatinghormone(FSH)duringthemenstrualcycle.Inadultwomen,Estradiollevelsaremeasuredintheevaluationoffertilityandmenstrualirregularities,andtomonitorovarianfollicularfunctionduringinductionofovulation.Inthefemale,Estradiolactsasagrowthhormonefortissueofthereproductiveorgans.ThedevelopmentofsecondarysexualcharacteristicsinwomenisdrivenbyEstradiol.Estradiolisinvolvedalsoinmenfertility.Estradiolregulatesthebonemaintenance.Post-menopausewomenexperienceanacceleratedlosofbonemassduetoarelativeEstrogendeficiency.
Estradiolaffectstheproductionofmultipleproteinsincludinglipoproteins,bindingproteins,andproteinsresponsIBLeforbloodclotting.Estrogenshavebeenfoundtohaveneuroprotectivefunction.
TheEstradiol,forhisactivities,isinvolvedinsometypesofcancersuchasbreastcancerandcanceroftheuterinelining.InadditionthereareseveralbenigngynaecologicconditionsthataredependentonEstrogensuchasendometriosis,leiomyomatauteri,anduterinebleeding.

Principlesoftheassay:Microtiterstripwellsareprecoatedwithanti-Estradiolantibodies(solid-phase).EstradiolinthesamplecompeteswithaddedhorseradishperoxidaselabelledEstradiol(enzyme-labelledantigen)forantibodybinding.Afterincubationabound/freeseparationisperformedbysolid-phasewashing.Theimmunecomplexformedbyenzyme-labelledantigenisvisualizedbyaddingTetramethylbenzidine(TMB)substratewhichgivesabluereactionproduct.TheintensityofthisproductisinverselyproportionaltotheamountofEstradiolinthesample.Sulphuricacidisaddedtostopthereaction.Thisproducesayellowendpointcolour.Absorptionat450nmisreadusinganELISAmicrowellplatereader.

StorageandStABIlity:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°Cinthedark.

LimitationsoftheTest:Sample(s),whicharecontaminatedmicrobiologically,shouldnotbeusedintheassay.Highlylipemicorhaemolysedspecimen(s)shouldsimilarlynotbeused.Itisimportantthatthetimeofreactionineachwellisheldconstantforreproducibleresults.Pipettingofsamplesshouldnotextendbeyondtenminutestoavoidassaydrift.Ifmorethanoneplateisused,itisrecommendedtorepeatthedoseresponsecurve.Additionofthesubstratesolutioninitiatesakineticreaction,whichisterminatedbytheadditionofthestopsolution.Therefore,theadditionofthesubstrateandthestoppingsolutionshouldbeaddedinthesamesequencetoeliminateanytimedeviationduringreaction.Platereadersmeasurevertically.Donottouchthebottomofthewells.Failuretoremoveadheringsolutionadequatelyintheaspirationordecantationwashstep(s)mayresultinpoorreplicationandspuriousresults.

ReferencesJoshi,U.M.etal.(1979)Steroids34(1),35Exley,D.andAbuknesha,R.(1978)FebsLetters91(2),162Ismail,A.A.,Niswender,G.D.andMidgley,A.R.(1972)J.Clin:Endocrin.Metab34,177-184Rajkowski,K.M.,CittanovaN.,Desfosses,B.andJayle,M.F.(1977)Steroids29(5),701-713Sadem,D.,Sela,E.andHexter,C.S.(1979)J.ofImmunologicalMethods28,125-131Wisdom,G.B.(1976)ClinChem.22(8),1243-1255

AdditionalInformation:

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Name17beta-ESTRADIOL
RelatedProductNames17β-Estradiol
MolecularWeight0.5
StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°Cinthedark.
IntendedUseResearchUseOnly