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Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg
  • Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg

Millipore/MAB5256 | Anti-Neurofilament 200 kDa Antibody, clone NE14/MAB5256/40 µg

價格: ￥4440.00 市場價: 7400.00
貨號: MAB5256
品牌: Millipore
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    • Description
      CatalogueNumberMAB5256
      BrandFamilyChemicon®
      TradeName
      • Chemicon
      DescriptionAnti-Neurofilament200kDaAntibody,cloneNE14
      BackgroundInformationNeurofilamentsareatypeofintermediatefilamentthatserveasmajorelementsoftheCytoskeletonsupportingtheaxoncytoplasm.Theyarethemostabundantfibrillarcomponentsoftheaxon,beingonaverage3-10timesmorefrequentthanaxonalmicrotubules.Neurofilaments(10nmindia.)arebuiltfromthreeintertwinedprotofibrilswhicharethemselvescomposedoftwotetramericprotofilamentcomplexsofmonomericproteins.Theneurofilamenttripletproteins(68/70,160,and200kDa)occurinboththecentralandperipheralnervoussystemandareusuallyneuronspecific.The68/70kDaNF-Lproteincanself-assembleintoafilamentousstructure,howeverthe160kDaNF-Mand200kDaNF-Hproteinsrequirethepresenceofthe68/70kDaNF-Lproteintoco-assemble.Neuromas,ganglioneuromas,gangliogliomas,ganglioneuroblastomasandneuroblastomasstainpositivelyforneurofilaments.Althoughtypicallyrestrictedtoneurons,neurofilamentshavebeendetectedinparagangliomasandadrenalandextra-adrenalpheochromocytomas.Carcinoids,neuroendocrinecarcinomasoftheskin,andoatcellcarcinomasofthelungalsoexpressneurofilaments.FormoreneurofilamentinformationseeNervousSystemCellTypeSpecificMarkerchartonlineundertheCHEMICONTechnicalSupportsection.
      ProductInformation
      FormatPurified
      PresentationPurifiedimmunoglobulin.Liquid.Buffer=0.02MPhosphatebuffer,0.25MNaClcontaining0.1%sodiumazide.
      StorageandShippingInformation
      StorageConditionsMaintainrefrigeratedat2-8°Cinundilutedaliquotsforupto6months.DONOTFREEZE
      Applications
      ApplicationDetectNeurofilament200kDausingthisAnti-Neurofilament200kDaAntibody,cloneNE14validatedforuseinIH.
      KeyApplications
      • Immunohistochemistry
      ApplicationNotesImmunohistochemistry:5-10μg/mL(Seebelowprotocol.)

      Optimalworkingdilutionsmustbedeterminedbyenduser.

      ImmunohistochemistryProtocolforAnti-Neurofilament200kD

      Idealspecimensareobtainedfromfrozensectionsfromshock-frozentissuesamples.Thefrozensectionsaredriedintheairandthenfixedwithacetoneat-15to-25°Cfor10min.Excessacetoneisallowedtoevaporateat15-25°C.Materialfixedinalcoholandembeddedinparaffincanalsobeused,see(Altmannsbergeretal.,1982).Theantibodyappearstoreactwithtissuefixedinformaldehydeforashorttime(10min)(Debusetal.,1983).Otherfixationconditionsmustbefirsttestedbytheinvestigator.

      Itisadvantageoustoblockunspecificbindingsitesbyoverlayingthesectionswithfetalcalfserumfor20-30minat15-25°C.Excessoffetalcalfserumisremovedbydecantingbeforeapplicationoftheantibodysolution.Cytocentrifugepreparationsofsinglecellsorcellsmearsarealsofixedinacetone.Thesepreparationsshould,however,notbedriedintheair.Instead,theexcessacetoneisremovedbybrieflywashinginphosphate-bufferedsaline(PBS).

      Furthertreatmentisthenasfollows:

      Overlaythepreparationwith10-20μLantibodysolutionandincubateinahumidchamberat37°Cfor1h.

      DiptheslidebrieflyinPBSandthenwash3timesinPBSfor3min(usefreshPBSeachtime)

      Wipethemarginsofthepreparationdryandoverlaythepreparationwith10-20μLofananti-mouseIg-FITCoranti-mouseIg-PODantibodyandallowtoincubatefor1hat37°Cinahumidchamber.

      Washtheslideasdescribedabove.

      Thepreparationmustnotbeallowedtodryoutduringanyofthesteps.

      Ifusinganindirectimmunofluorescencetechnique,thepreparationshouldbeoverlaidwithasuitableembeddingmedium(e.g.Moviol,Hoechst)andexaminedunderthefluorescencemicroscope.IfaPOD-conjugatehasbeenusedasthesecondaryantibody,thepreparationshouldbeoverlaidwithasubstratesolution(seebelow)andincubatedat15-25°CuntilaclearlyvisIBLered-browncolordevelops.Anegativecontrol(e.g.onlythesecondaryantibody)shouldremainunchangedincolorduringthisincubationperiod.Subsequently,thesubstrateiswashedoffwithPBSandthepreparationisstained,ifdesired,withhemalumstainforabout1min.ThehemalumsolutioniswashedoffwithPBS,thepreparationisembeddedandexamined.

      Substratesolutions:

      Aminoethyl-carbazole:

      Dissolve2mg3-amino-9-ethylcarbazolewith1.2mLdimethylsulfoxideandadd28.8mL0.05MTris-HCl,pH7.3,and20μL3%H2O2(w/v).Preparesolutionfreshlyeachday.

      Diaminobenzidine:

      Dissolve25mg3,3"-diaminobenzidinewith50mL0.05MTris-HCl,pH7.3,andadd40μL3%H2O2(w/v).Preparesolutionfreshlyeachday.
      BIOLOGicalInformation
      ImmunogenPurifiedneurofilamentpolypeptides(Debusetal.,1983).
      CloneNE14
      ConcentrationPleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
      HostMouse
      SpecificityTheantibodyreactswithneurofilament200kD.
      IsotypeIgG1
      SpeciesReactivity
      • Human
      • Pig
      • Rat
      AntibodyTypeMonoclonalAntibody
      EntrezGeneNumber
      GeneSymbol
      • NEFH
      • NFH
      • NF-H
      • KIAA0845
      UniProtNumber
      UniProtSummaryFUNCTION:SwissProt:P12036#Neurofilamentsusuallycontainthreeintermediatefilamentproteins:L,M,andHwhichareinvolvedinthemaintenanceofneuronalcaliber.NF-HhasanimportantfunctioninmatureaxonsthatisnotsubservedbythetwosmallerNFproteins.
      SIZE:1026aminoacids;112480Da
      PTM:ThereareanumberofrepeatsofthetripeptideK-S-P,NFHisphosphorylatedonanumberoftheserinesinthismotif.ItisthoughtthatphosphorylationofNFHresultsintheformationofinterfilamentcrossbridgesthatareimportantinthemaintenanceofaxonalcaliber.&Phosphorylationseemstoplayamajorroleinthefunctioningofthelargerneurofilamentpolypeptides(NF-MandNF-H),thelevelsofphosphorylationbeingaltereddevelopmentallyandcoincidentwithachangeintheneurofilamentfunction.
      DISEASE:SwissProt:P12036#DefectsinNEFHareacauseofsusceptibilitytoamyotrophiclateralsclerosis(ALS)[MIM:105400].ALSisaneurodegenerativedisorderaffectingupperandlowermotorneurons,andresultinginfatalparalysis.Sensoryabnormalitiesareabsent.Deathusuallyoccurswithin2to5years.Theetiologyislikelytobemultifactorial,involvingbothgeneticandenvironmentalfactors.
      SIMILARITY:SwissProt:P12036##Belongstotheintermediatefilamentfamily.
      PhysicochemicalInformation
      Dimensions
      MaterialsInformation
      MaterialsInformation
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