Description:ProductName:LH#ofSamples:1x96AssaysIntendedUse:Immunoenzymaticcolorimetricmethod(ELISA)forquantitativedeterminationofLHinserum.Introduction:Luteinizinghormone(LH)isaglycoproteinconsistingoftwosubunitswithamolecularmassof30,000daltons.Theα-subunitissimilartootherpituitaryhormones[folliclestimulatinghormone(FSH),thyroidstimulatinghormone(TSH)andchorionicgonadotropin(HCG)]whiletheβ-subunitisunique.Theβ-subunitconferstheBIOLOGicalactivitytothemolecule.Theα-subunitconsistsof89aminoacidresidueswhiletheβ-subunitcontains129aminoacids.Thecarbohydratecontentisbetween15%and30%.Theclinicalusefulnessofthemeasurementofluteinizinghormone(LH)inascertainingthehomeostasisoffertilityregulationviathehypothalamic-pituitary-gonadalaxishasbeenwellestablished(1,2).Inaddition,theadventofinvitrofertilization(IVF)technologytoovercomeinfertilityassociatedproblemshasprovidedtheimpetusforrapidimprovementinLHassaymethodologyfromthetechnicallydemandingbioassaytotheprocedurallysimpleandrapidimmunoenzymometricassays.Principlesoftheassay:Inthismethod,LHstandards,patientspecimensand/orcontrols(containingthenativeantigen)arefirstaddedtostreptavidincoatedwells.Biotinylatedmonoclonalandenzymelabeledantibodiesareaddedandthereactantsmixed:theseantibodieshavehighaffinityandspecificityandaredirectedagainstdistinctanddifferentepitopesofLH.ReactionbetweenthevariousLHantibodiesandnativeLHoccursinthemicrowellswithoutcompetitionorsterichindranceformingasolublesandwichcomplex.Theinteractionisillustratedbythefollowingequation:kaEnzAb+AgLH+BtnAb(m)?EnzAb-AgLH-BtnAb(m)k-aBtnAb(m)=BiotinylatedMonoclonalAntibody(ExcessQuantity)AgLH=NativeAntigen(VariableQuantity)EnzAb(p)=EnzymelabeledAntibody(ExcessQuantity)EnzAb(p)-AgLH-BtnAb(m)=Antigen-AntibodiesSandwichComplexka=RateConstantofAssociationk-a=RateConstantofDissociationSimultaneously,thecomplexisdepositedtothewellthroughthehighaffinityreactionofstreptavidinandbiotinylatedantibody.Thisinteractionillustratedbelow:EnzAb-AgLH-BtnAb(m)+StreptavidinC.W.?????ImmobilizedcomplexStreptavidinC.W.=Streptavidinimmobolizedonwell.Immobilizedcomplex=Antibodies-Antigensandwichbound.Afterequilibriumisattained,theantibody-boundfractionisseparatedfromunboundantigenbydecantationoraspiration.Theenzymeactivityintheantibody-boundfractionisdirectlyproportionaltothenativeantigenconcentration.Theactivityoftheenzymepresentonthesurfaceofthewellquantitatedbyreactionwithasuitablesubstratetoproducecolour.Byutilizingseveraldifferentcalibratorsofknownantigenvalues,adoseresponsecurvecanbegeneratedfromwhichtheantigenconcentrationofanunknowncanbeascertainedStorageandStABIlity:Thereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat+2...+8°Cinthedark.Openedreagentsarestablefor60dayswhenstoredat+2...+8°C.LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.LHissuppressedbyestrogenbutinwomantakingoralcontraceptivesthelevelmaybelowornormal.Excessivedietingandweightlossmayleadtolowgonadotropinconcentrations.Luteinizinghormoneisdependentupondiversefactorsotherthanpituitaryhomeostasis.Thus,thedeterminationaloneisnotsufficienttoassessclinicalstatusReferencesKosasaT.S.,"MeasurementofHumanLuteinizingHormone."JournalofReproductiveMedicine,26(1981)pg.201-6.2.DanzerH.,BraunsteinG.D.,etal.,"MaternalSerumHumanChorionicGonadotropicConcentrationsandFetalSexPredictions."FertilityandSterility,34(1980)pg.336-40.3.BraunsteinG.D.,etat.,"SerumHumanLuteinizingHormoneLevelsthroughNormalPregnancy",AmericanJournalofObstetricsandGynecology,126(1976)pg.678-81.4.GoldsteinD.P.,andKosasaT.S.,"TheSubunitRADIoimmunoassayforLHClinicalApplication."Gynecology,6(1975)pg.145-84.5.BatzerF.,"HormonalEvaluationofEarlyPregnancy",FertilityandSterility,34(1980)pg.1-12.6.Braunstein,G.D.,etal.,"First-TrimesterLuteinizingHormoneMeasurementsasanAidtotheDiagnosisofEarlyPregnancyDisorders”,AmericanJournalofObstetricsandGynecology,131(1978)pg.25-32.7.LentonE.,NealL.andSulaimanR.,"PlasmaConcentrationsofHumanGonadotropinfromthetimeofImplantationuntiltheSecondWeekofPregnancy",FertilityandSterility,37(1982)pg.773-78.AdditionalInformation:NameLHRelatedProductNamesLHMolecularWeight0.5StorageThereagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat+2...+8°Cinthedark.Openedreagentsarestablefor60dayswhenstoredat+2...+8°C.IntendedUseResearchUseOnly