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

價格: ￥5052.00 市場價: 8420.00

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貨號: AG105

品牌: Millipore

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Description
CatalogueNumber	AG105
TradeName	BLACK-GOLD
Description	BlackGoldIIMyelinStainingKit
Overview	Black-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
Presentation	SeeComponents

StorageandShippingInformation
StorageConditions	Thekitcanbestoredatroomtemperature.Ideally,theBlack-GoldIIpowdershouldbekeptinadesiccator,becauseofitshygroscopicnature.Thedrypowder,ifunopenedandstoredproperly,isuseableforuptooneyearfromdateofreceipt.OnceresUSPendedtheBlack-GoldIIstainingsolutionshouldbestoredinthedarkat4°Candisgoodforupto3months. TOXICITY:Althoughthecompoundappearstobeoflowtoxicity,ithasnotbeenextensivelyevaluatedandthereforeroutinelaboratorycautionshouldbeexercised.Notintendedforhumanconsumption.

StorageandShippingInformation

StorageConditions

Thekitcanbestoredatroomtemperature.Ideally,theBlack-GoldIIpowdershouldbekeptinadesiccator,becauseofitshygroscopicnature.Thedrypowder,ifunopenedandstoredproperly,isuseableforuptooneyearfromdateofreceipt.OnceresUSPendedtheBlack-GoldIIstainingsolutionshouldbestoredinthedarkat4°Candisgoodforupto3months.

TOXICITY:Althoughthecompoundappearstobeoflowtoxicity,ithasnotbeenextensivelyevaluatedandthereforeroutinelaboratorycautionshouldbeexercised.Notintendedforhumanconsumption.

Applications
KeyApplications	Immunohistochemistry
ApplicationNotes	TissueFixation: 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.

Applications

KeyApplications

Immunohistochemistry

ApplicationNotes

TissueFixation:
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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