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    當前位置 ： Millipore >>> Millipore/AG325 | Fluoro-Jade? C/AG325/50 mg
    Millipore/AG325 | Fluoro-Jade? C/AG325/50 mg
    • Millipore/AG325 | Fluoro-Jade? C/AG325/50 mg

    Millipore/AG325 | Fluoro-Jade? C/AG325/50 mg

    價格: 試用 市場價: 0.00
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    貨號: AG325
    品牌: Millipore
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      • Description
        CatalogueNumberAG325
        BrandFamilyChemicon®
        TradeName
        • Fluoro-Jade
        • Chemicon
        DescriptionFluoro-Jade?C
        OverviewFluoro-Jade?C,likeitspredecessors,Fluoro-Jade?andFluoro-Jade?B,werefoundtostainalldegeneratingneurons,regardlessofspecificinsultormechanismofcelldeath.Therefore,thepatternsofneuronaldegenerationseenfollowingexposuretoeithertheglutamateagoNIST,kainicacid,ortheinhibitorofmitochondrialrespiration,3-NPA,werethesameforalloftheFluoro-Jadeadyes.However,therewasaqualitativedifferenceinthestainingcharacteristicsofthethreeFluorochromes.Specifically,Fluoro-Jade?Cexhibitedthegreatestsignaltobackgroundratio,aswellasthehighestresolution.Thistranslatestoastainofmaximalcontrastandaffinityfordegeneratingneurons.Thismakesitidealforlocalizingnotonlydegeneratingnervecellbodies,butalsodistaldendrites,axonsandterminals.ThedyeishighlyresistanttofADIngandiscompatIBLewithvirtuallyallhistologicalprocessingandstainingprotocols.TriplelabelingcanbeaccomplishedbystainingdegeneratingneuronswithFluoro-Jade?C,cellnucleiwithDAPIandactivatedastrocyteswithGFAPimmunofluorescence.



        APPEARANCE:Coffeebrowntobrickredpowder.

        MOLECULARWEIGHT:823

        EXCITATIONPEAK:485nm

        EMISSIONPEAK:525nm

        FILTERSYSTEM:Fluorescein/FITC

        SOLUBILITY:Highlysolubleinwaterandbases,moderatelysolubleinalcoholandweakacids.

        TOXICITY:Althoughthecompoundappearstobeoflowtoxicity,ithasnotbeenextensivelyevaluatedandthereforeroutinelaboratorycautionshouldbeexercised.Notintendedforhumanconsumption.
        BackgroundInformationFluoro-Jade?C,likeitspredecessors,Fluoro-Jade?andFluoro-Jade?B,werefoundtostainalldegeneratingneurons,regardlessofspecificinsultormechanismofcelldeath.Therefore,thepatternsofneuronaldegenerationseenfollowingexposuretoeithertheglutamateagonist,kainicacid,ortheinhibitorofmitochondrialrespiration,3-NPA,werethesameforalloftheFluoro-Jade?dyes.However,therewasaqualitativedifferenceinthestainingcharacteristicsofthethreefluorochromes.Specifically,Fluoro-Jade?Cexhibitedthegreatestsignaltobackgroundratio,aswellasthehighestresolution.Thistranslatestoastainofmaximalcontrastandaffinityfordegeneratingneurons.Thismakesitidealforlocalizingnotonlydegeneratingnervecellbodies,butalsodistaldendrites,axonsandterminals.Thedyeishighlyresistanttofadingandiscompatiblewithvirtuallyallhistologicalprocessingandstainingprotocols.TriplelabelingcanbeaccomplishedbystainingdegeneratingneuronswithFluoro-Jade?C,cellnucleiwithDAPIandactivatedastrocyteswithGFAPimmunofluorescence.Fluoro-Jade?isaregisteredtrademarkofHisto-Chem,Inc.
        ProductInformation
        StorageandShippingInformation
        StorageConditionsThelyophilizedpowdershouldbestoredwellsealedatroomtemperature,preferableinadesiccator(duetoitshygroscopicnature)foruptooneyearfromdateofreceipt.Theliquidstocksolution(0.01%)indistilledwatercanbestoredat2-8°Cforupto3months.The.0002-.0001%workingsolutionin0.1%aceticacidshouldbeusedwithin4hrsofpreparation.
        Applications
        KeyApplications
        • Immunohistochemistry
        ApplicationNotesSUGGESTEDPROTOCOLFORUSINGFLUORO-JADE?C

        Processing:Halfofeachgroupofbrainswereparaffinembeddedandcutonarotarymicrotomewhiletheremainderwerecutonafreezingslidingmicrotome.Paraffinsectionswere10uminthicknesswhilefrozensectionswerecutatathicknessof25um.Priortostaining,sectionsweremountedfromdistilledwaterontogelledslides.Gelatincoatedslideswerepreparedbyimmersionina60degreeCsolutionof1%pigskingelatin(Sigma;typeA,300Bloom)andthenovendriedovernightatthesametemperature.Thesectionsweremountedontotheslidesfromdistilledwaterandthenairdriedforatleast30minonaslidewarmerat50degreesC.Slidesbearingfrozencuttissuesectionswerefirstimmersedinabasicalcoholsolutionconsistingof1%sodiumhydroxidein80%ethanolfor5min.Theywerethenrinsedfor2minin70%ethanol,for2minindistilledwater,andthenincubatedin0.06%potassiumpermanganatesolutionfor10min.Followinga1-2minwaterrinse,theslideswerethentransferredfor10mintoa0.0001%solutionofFluoro-Jade?Cdissolvedin0.1%aceticacidvehicle.Theproperdilutionwasaccomplishedbyfirstmakinga0.01%stocksolutionofthedyeindistilledwaterandthenadding1mLofthestocksolutionto99mLof0.1%aceticacidvehicle.Theworkingsolutionwasusedwithin2hofpreparation.Thestocksolution,whenrefrigerated,canbekeptforlongperiodsbutshouldbediscardedifthesolutionbecomescloudy.Theslideswerethenrinsedthroughthreechangesofdistilledwaterfor1minperchange.Excesswaterwasdrainedontoapapertowel,andtheslideswerethenairdriedonaslidewarmerat50degreesCforatleast5min.Theairdriedslideswerethenclearedinxyleneforatleast1minandthencoverslippedwithDPX(FlukaorSigma)nonfluorescentmountingmedia.Polarcoverslippingmedia,suchasthosethatcontainwater,alcoholorglycerolwereneverused.Forcomparativepurposes,someslideswerestainedwithFluoro-Jade?Baccordingtothepreviouslydescribedprocedure.Whenworkingwithparaffinprocessedtissue,thesectionsarefirstdeparaffinizedthroughtwo10minchangesofxyleneandthenthesectionsarerehydratedthroughagraduatedalcoholseries,omittingthebasicalcoholsolution.Onceindistilledwater,thesectionsaretransferredtothepotassiumpermanganatesolutionatwhichpointthestainingprocedureisidenticaltothatdescribedforfrozensections.Multiplelabeling:Fluoro-Jade?CcanreadilybecombinedwithotherfluorescentMarkers.Multiplelabelingwasachievedusinganti-glialfibrillaryacidicprotein(GFAP)immunocytochemistrytolabelactivatedastrocyteswhileusingDAPItolabelnuclearDNA.Incorporating4",6-diamidino-2-phenylindole(DAPI;Sigma,St.LouisMO)asafluorescentnuclearstainisaccomplishedbysimplyincorporating0.0001%intotheFluoro-Jade?Cstainingsolution.Thisisaccomplishedbytheadditionof1mlof0.01%DAPIstocksolutionto99mlof0.1%aceticacid.Fluoro-Jade?CwasalsocombinedwithimmunofluorescentlabelingofGFAPaccordingtothefollowingprocedure.Loosefrozentissuesectionswereincubatedinapredilutedsolutionofanti-GFAP(ChemiconhasanumberofdifferentantibodiestoGFAP)atabout5degreesCintherefrigeratorfor1-3days.Itshouldbementionedthatalthoughinthisstudyallimmunocytochemistrywasperformedonfrozensections,themethodsarefullycompatiblewithparaffinprocessedtissueaswell.Sectionswererinsedintwochangesofbufferedsalinefor10mineachandthentransferredtoatetramethylrhodamineisothiocynate(TRITC)labeledsecondaryantibody(ChemiconhasanumberofdifferentTRITClabeledsecondaryantibodies),diluted1:100inbufferedsaline,for1hatroomtemperature.Sectionswererinsedintwochangesofbufferedsalinefor10mineachandthenthesectionsweremountedontogelledslidesfromdistilledwaterandairdriedonaslidewarmerat50degreesCfor30min.TocombinewithFluoro-Jade?C,theslidemountedsectionswererehydratedfor2minindistilledwaterandthentransferredtothe0.06%potassiumpermanganatesolutionfor10min.Itisworthmentioningthattheincubationtimeinpotassiumpermanganatemayneedtobereducedwhenco-localizingthoseantigenicepitopessusceptibletochemicaloxidation.Theslideswerethenrinsedfor2minindistilledwater,transferredtotheFluoro-Jade?Cworkingsolutionfor10minandthenrinsed,airdehydrated,xyleneclearedandcoverslippedwithDPX,aspreviouslydescribed.ThebluenuclearlabelconferredbyDAPIisvisualizedviaultravioletlightexcitation,whiletheredTRITClabeledantibodyisvisualizedbygreenlightexcitation.
        BIOLOGicalInformation
        PuritySilicaTLC(acetanitrile/water,6/4)revealedthepresenceoftwofluorescentspots,presumablycorrespondingtothedisulphatehomologues.Thepresenceofprecursorsorfreefluoresceinwasnotdetected.
        PhysicochemicalInformation
        Dimensions
        MaterialsInformation
        MaterialsInformation
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