Overview

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Ex/Em(nm)	620/None
MW	N/A
CAS#	N/A
Solvent	N/A
Storage	F/D/L
Category	SmallMoleculeDetection MetalIons
Related	BiochemicalAssays

Zincisanessentialtracemineralelementthatplaysanimportantroleinanumberofbiologicalprocesses.Itisanessentialfactorrequiredformanyenzymes,proteinstructures,andcontrolofgeneticexpression.Zincstatusalsoaffectsbasicprocessesofcelldivision,growth,differentiation,development,andaging.Clinicalsignsofzincdeficiencyincludeacrodermatitis,lowimmunity,diarrhea,poorhealing,stunting,hypogonadism,fetalgrowthfailure,teratologyandabortion.Simple,directandautomation-readyproceduresformeasuringarehighlydesirableinresearchanddrugdiscovery.AATBioquest"sAmplite™ColorimetricZincQuantitationKitprovidesasimplemethodfordetectingzincconcentrationinbiologicalsamplesusingourproprietaryZin-620™,inwhichZincbindstotheprobewiththeenhancedabsorptionaround620nm.TheZincprobeexhibitsalargeincreasein620nmabsorptioninresponsetoZn2+(>100folds).OurkitformulationhasenhancedZn2+-specificitywithlittleresponsestoothermetals,e.g.,Ca2+andMg2+.TheassaycanbeusedwithBIOLOGicalsamplessuchasserum,plasma,andurinewithdetectionsensitivityat1µM.OurAmplite™ColorimetricZincQuantitationKit(#19000)isevenmoresensitive,andcanbeusedfordetectingaslowas0.1uMZnion.

Thisprotocolonlyprovidesaguideline,andshouldbemodifiedaccordingtoyourspecificneeds.

1.      ZnCl₂standardsandtestsamplepreparations:

1.1         Add10μLof100mMZnCl₂Standardsolution(ComponentC)into990µLAssayBuffer(Component

           B)toget1mMZnCl₂standardsolution. 

 

1.2         Add100μLof1mMZincStandardsolution(ComponentC)to990µLAssayBuffer(ComponentB)to

           get100µMZnCl₂standardsolution.

 

1.3         Take300μLof100µMZnCl₂standardsolution(fromStep1.2)toperform1:2serialdilutionstoget

           50,25,12.5,6.25,3.13,1.56and0µMseriallydilutedZnCl₂standards.

 

1.4         Dilutethetestsampleto5-100µMrangewithAssayBuffer(ComponentB).

 

1.5         Add50μLofZnCl₂standards,dilutedsampleandcontrolintoeachwellasshowninTable1and2.

Table1.LayoutofZnCl₂standardsandtestsamplesinaclearbottom96-wellmicroplate

Note:Zn=ZincStandards,BL=BlankControl,TS=TestSamples.

 

Table2.Reagentcompositionforeachwell

ZincStandard	BlankControl	TestSample
Serialdilutions*:50μL	AssayBuffer:50μL	50μL

*Note:AddtheseriallydilutedZnCl₂standardsfrom100µMto1.56µMintowellsfromZn1toZn7induplicate.

 

2.      RunZincassay:

2.1          Add25μLof Zn-620™(ComponentA)into5mLAssayBuffer(ComponentB)tomakeZnassay

mixture.Add50μLZnassaymixturetoeachwellofZnCl₂standard,blankcontrol,andtestsamples(seeStep1.5,Table1and2)tomakethetotalZnCl₂assayvolumeof100µL/well.

Note: Fora384-wellplate,add25μLofsamplesand25μLofassayreactionmixtureintoeachwell.

2.2         Incubatethereactionfor5-10minutesatroomtemperature,protectedfromlight.

2.3         Monitortheabsorbanceincreasewithaabsorbanceplatereaderat610nmand480nm.

References&Citations

CitationExplorer

Afteroxidation,zincnanoparticleslosetheirABIlitytoenhanceresponsestoodorants
Authors:SamanthaHagerty,YasmineDaniels,MelissaSingletary,OlegPustovyy,LudmilaGloba,WilliamAMacCrehan,ShinMuramoto,GheorgheStan,JuneWLau,EdwardEMorrison
Journal:BioMetals(2016):1--14

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