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Millipore/AG105 | Black Gold II Myelin Staining Kit/AG105/1
  • Millipore/AG105 | Black Gold II Myelin Staining Kit/AG105/1

Millipore/AG105 | Black Gold II Myelin Staining Kit/AG105/1

价格: ¥5052.00 市场价: 8420.00
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货号: AG105
品牌: Millipore
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    • Description
      CatalogueNumberAG105
      TradeName
      • BLACK-GOLD
      DescriptionBlackGoldIIMyelinStainingKit
      OverviewBlack-GoldIIisanaurohalophosphatecomplexwhichstainsspecificallyformyelinwithinthecentralnervoussystem.Black-GoldIIstainingprovideshighresolution,highcontrast,ashorthistochemicalprocessingtime,andhighreproducibility.Black-GoldIIisanewandimprovedversionofitspredecessor,Black-Gold(Cat#AG390).TheadvantagesofMillipore’sBlack-GoldIIMyelinStainingKitoverBlackGoldIarethatitismorereADIlysoluble,canbeutilizedatahigherconcentration,producesamoreuniformandconsistentstaining,takeslesstimetostain,anddoesnotrequirepost-stainingintensificationprocedures.

      TheuseofBlack-GoldIIistailoredtostudiesusingformalin-fixed,non-solventprocessedtissue.Thetechniquestainslargemyelinatedtractsdarkred-brown,whiletheindividualmyelinatedfibersappearblack.Thisnoveltracercanbeusedtolocalizebothnormalandpathologicalmyelin.Black-GoldIIcandemonstrateandcharacterizespecificmyelinchangesassociatedwithexposuretodiverseneurotoxicantsincludingkainicacid,methamphetamine,acrylamide,domoicacid,3-nitropropionicacid,Fluoro-Goldandisoniazid.

      TheBlack-GoldIIMyelinStainingKitalsocomeswithsodiumthiosulfateandcresylviolet.ThesodiumthiosulfatesolutionisrequiredforextractingexcessBlack-GoldIIstain.WhereasthecresylvioletisaNisslstainthatcolorscellbodiesabrilliantbluishhueandisusedasacounterstainwithBlack-GoldII.


      BlackGoldIIPurity:Nodetectableamountofuncomplexedgoldwasfound.

      BlackGoldIIIllumination:Brightfieldordarkfield.

      BlackGoldIISolubility:Freelysolubleinwater,saline,ordiluteacids.
      ProductInformation
      Components
      • Black-GoldII:150mgdrypowder,yellowincolor.
      • CresylVioletStain:150mgdrypowder,purplish-blackincolor.
      • 5%SodiumThiosulfate:10ml
      PresentationSeeComponents
      StorageandShippingInformation
      StorageConditionsThekitcanbestoredatroomtemperature.Ideally,theBlack-GoldIIpowdershouldbekeptinadesiccator,becauseofitshygroscopicnature.Thedrypowder,ifunopenedandstoredproperly,isuseableforuptooneyearfromdateofreceipt.OnceresUSPendedtheBlack-GoldIIstainingsolutionshouldbestoredinthedarkat4°Candisgoodforupto3months.

      TOXICITY:Althoughthecompoundappearstobeoflowtoxicity,ithasnotbeenextensivelyevaluatedandthereforeroutinelaboratorycautionshouldbeexercised.Notintendedforhumanconsumption.
      Applications
      KeyApplications
      • Immunohistochemistry
      ApplicationNotesTissueFixation:
      Tissuecanbefixedwitheitherformalinor4%PFA.Typicallyanimalsareperfusedwithneutralphosphatebuffered10%formalin(or4%formaldehyde)viatheascendingaorta.Thebrainsarethenremovedandfixedovernightinthesamefixativesolution.Longerfixationtimesaregenerallynotrecommended.Forcryosections,thebrainsmaybeprotectedthroughtreatmentwithsucrosesolution(20-30percent).Subsequentlyfrozenbraintissueiscutonafreezingslidingmicrotome.Note:Black-GoldIIwillnotstainparaffinembeddedorunfixedtissue.

      Sectioning:
      Eitherfrozenorfixed,non-frozenvibratomesectionscanbeusedandshouldbecutatathicknessof20-40μm.Thesectionsarethentypicallymountedonpositivelychargedslides.Thesectionscanalsobestainedloose,althoughthesectionsareeasiertohandlewhenmountedonslides.

      Solutions:
      Black-GoldIIpowdershouldberesuspendedwithsalinesolution(0.9%NaCl)toafinalconcentrationof0.3%.Simplyaddthe150mgofBlack-GoldIIpowderinto50mlsof0.9%NaClsolutionandmixtoresuspend.OnceresuspendedtheBlack-GoldIIstainingsolutionshouldbestoredinthedarkat4°Candisgoodforupto3months.
      CresylVioletpowdershouldberesuspendedtoafinalconcentrationof0.1%usingamixtureofMilliQwaterandglacialaceticacid(3µlofacidpermlofwater).Onceresuspendedthecresylvioletsolutionshouldbestoredat4°Candisgoodforupto6months.
      Thesodiumthiosulfateshouldbedilutedto1%usingMilliQwaterpriortouse.

      Staining:
      Pre-heat0.3%Black-GoldIIand1%sodiumthiosulfatesolutionsto60°C.RehydratetissuesectionsinMilliQwaterfor2minutesthentransfertopre-warmedBlack-GoldIIsolution.IncubateslidesinBlack-GoldIIsolutionat60°Cfor12minutes.Atthispointtheslidesshouldbemonitoredat2-3minuteintervalstodeterminetheextentoflabeling.Stainingiscompletewhenthefinestmyelinatedfibersarestainingdarkredtoblack.Theappearanceofalavenderbackgroundcolorindicatesoverstainingandtheprocessshouldbestopped.Note:Theexactstainingtimewillvarydependingonsectionthickness,fixingconditions,typeoftissue,andageofstainsolution.

      Thesectionsarethenrinsed2XinMilliQwaterfor2minuteseach.Sodiumthiosulfatesolution(1%)isthenaddedandincubatedfor3minutesat60°C.Thesectionsarethenrinsed3XwithMilliQwaterfor2minuteseach.Cresylvioletsolutionisthenaddedandincubatedfor3minutesatroomtemperature.Thesectionsarethenrinsed3XwithMilliQwaterfor2minuteseach.

      Post-stainingsteps:
      Finally,dehydratetheslidesusingaseriesofgradatedalcohols.Thecresylstainwillleachfromthetissueatthistimeandtheextentofstainingcanbecontrolledbyhowlongtheseincubationslast.Thedehydratedsectionsarethenclearedinhistoclearorxylenefor2minutesandthencoverslippedwithmountingmedia.


      FrequentlyAskedQuestions(FAQs):

      1)ItappearsthatMyelinimpregnationisincompletewithBlack-GoldII?
      Answer:Returnslidesto60°CBlack-GoldIIsolutionandmonitorevery2-3minutesundermicroscopeuntilfineparallelfibersoflayer1cortexarevisIBLe.

      2)Backgroundhaslavendercolor?
      Answer:Sectionhasbeenoverstainedandstainingtimeshouldbereduced.

      3)Backgroundstainingoccursbeforeimpregnationofallfinemyelinatedfibers?
      Answer:Tissuewasprobablyover-fixed.Trytoavoidleavingbraininfixative,orsectionsinbuffer,forlongerthanonemonth.

      4)WhataretheadvantagesoverothermyelinstainslikeLuxolfastblue,orSudanBlack?
      Answer:Betterresolutionandcontrast.Faster.

      5)DoothercounterstainsworkwithBlack-GoldII?
      Answer:Yes,mostconventionalNisslstains(includingblue,red,andgreen)canbeusedasacounterstain.Additionally,thecounterstaincanbeomittedifonlymyelinstainingisdesired.
      BIOLOGicalInformation
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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