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

價格: ￥4440.00 市場價: 7400.00

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貨號: MAB5256

品牌: Millipore

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Description
CatalogueNumber	MAB5256
BrandFamily	Chemicon®
TradeName	Chemicon
Description	Anti-Neurofilament200kDaAntibody,cloneNE14
BackgroundInformation	NeurofilamentsareatypeofintermediatefilamentthatserveasmajorelementsoftheCytoskeletonsupportingtheaxoncytoplasm.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
Format	Purified
Presentation	Purifiedimmunoglobulin.Liquid.Buffer=0.02MPhosphatebuffer,0.25MNaClcontaining0.1%sodiumazide.

StorageandShippingInformation
StorageConditions	Maintainrefrigeratedat2-8°Cinundilutedaliquotsforupto6months.DONOTFREEZE

Applications
Application	DetectNeurofilament200kDausingthisAnti-Neurofilament200kDaAntibody,cloneNE14validatedforuseinIH.
KeyApplications	Immunohistochemistry
ApplicationNotes	Immunohistochemistry: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
Immunogen	Purifiedneurofilamentpolypeptides(Debusetal.,1983).
Clone	NE14
Concentration	PleaserefertotheCertificateofAnalysisforthelot-specificconcentration.
Host	Mouse
Specificity	Theantibodyreactswithneurofilament200kD.
Isotype	IgG1
SpeciesReactivity	Human Pig Rat
AntibodyType	MonoclonalAntibody
EntrezGeneNumber	NM_021076.2
GeneSymbol	NEFH NFH NF-H KIAA0845
UniProtNumber	P12036
UniProtSummary	FUNCTION: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.

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

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