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IBL/IGF-II ELISA/MD58041/

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Kitsize12x8MethodELISAIncubationtime1x2h,1x30min,1x30minStandardrange0,016-3.609µg/LSpecimen/Volumes5µLSerum,PlasmaSubstrate/isotopeTMB,450nmRegulatoryStatus:EU:CEDetailsfor: IGF-IIELISATheIGF-IIELISAiscalibratedagainsttheInternationalStandard:WHONIBSC96/538.ThestandardsoftheELISAarehumanIGF-IIinconcentrations0.45;1.5;3;5.63and9ng/ml,respectivelytheassayrangecovers–atrecommendednormalsampledilution-therangeto2400ng/ml.Byvaryingthesampledilutionthiscanbeadaptedtothespecialindividualrequirements.SensitivityTheanalyticalsensitivityoftheELISAyields0.02ng/ml(2SDofzerostandardin20folddetermination).TheInter-andIntra-Assayvariationcoefficientsarelessthan7.2%and6.6%respectively.INTENDEDUSEScientificinvestigationsinthefieldofneonatalhypertrophy(IGF-IIisafoetalgrowthfactor)andmalignancies(IGF-IIisamonogenicgrowthfactor).IGF-IIseemstobeofuseindifferentialdiagnosticsofmalignancies.Thus,itispossIBLetodifferentiatebyIGF-IIbetweenadrenocorticalcarcinomasandadenomas.FurthertumorstaginganddifferentiationbetweenhyperplasiaandcarcinomacanbeimprovedbyIGF-IImeasurementsinprostatetumors.TheIGF-Systemseemtobeofrelevanceinneurodegenerationaswell,e.g.Alzheimer´sandParkinson’sdiseases.Theinsulin-likegrowthfactors(IGF)-Iand–IIplayapivotalroleintheregulationofproliferationanddifferentiationofseveraltissuetypes.IGF-IalsocalledSomatomedinChasamolecularweightof7.469kDa.ItsexpressionismainlyregulatedbyGrowthHormoneandnutrition.ButseveralhormonesandpeptidefactorsareknowntoinfluenceIGF-IIsynthesisindifferenttissues.BioavailABIlityoftheIGFsisregulatedbyspecificbindingproteins(IGFBP).BesidethehighaffinityInsulin-likeGrowthFactorBindingProteins1-6,IGFsarealsoboundbeIGFBP-relatedProteins.ThesebindingproteinsbindIGF-IandIGF-IIwiththesameaffinityorpreferIGF-II.DirectmeasurementofIGFsinserumsampleswithoutpretreatmentresultsinfalsevaluesbecauseoftheextremelyslowdissociationoftheIGF/IGFBPcomplexesduringtheassayincubationonlyapartoftheIGF-IIinthespecimencanbindtotheantibodiesandbedetected.Therefore,varioustechniqueswereappliedtophysicallyseparateIGF-IIfromitsbindingproteinsbeforemeasurement,including(a)sizeexclusionchromatographyunderacidicconditions,(b)solid-phaseextractionand(c)acid-ethanolextraction.Thesetechniques,however,areeitherinconvenientortime-consumingorgiveincompleteandnot-reproduciblerecoveries.Thisassayiseasy,fastandresultsdonotdependonthebindingproteinconcentrationofthesample.ItisbasedonthehighspecificityoftheemployedantibodiesforIGF-II.Thereisvirtuallynocross-reactivitywithIGF-I.ThisallowstheseparationofIGF-IIfromthebindingproteinsbyacidificationandblockingofthefreebindingproteinswithIGF-I.Thus,theendogenousIGF-IIisfreeinsolution.ForconcretedatapleaseconsulttheInstructionforUseinthedownloadboxontherightside.

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