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Millipore/NS170 | Phosphorylated Neurofilament (pNF-H) Sandwich ELISA Kit/NS170/1 kit96 wells

價格: ￥6972.00 市場價: 11620.00

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貨號: NS170

品牌: Millipore

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Description
CatalogueNumber	NS170
BrandFamily	Chemicon®
TradeName	Chemicon
Description	PhosphorylatedNeurofilament(pNF-H)SandwichELISAKit
BackgroundInformation	TheChemiconPhosphorylatedNeurofilamentH(pNF-H)kitisasandwichenzymeimmunoassay(EIA),whichmeasurespNF-H.ThekitwillmeasurepNF-Hfromavarietyofmammalianspeciesincludinghuman,rat,mouse,bovine,porcine,andfeline. Neurofilamentsarethe10nmdiameterfilamentswhichmakeupthemajorpartoftheCytoskeletonofneuronsandareparticularlyconcentratedinlargerdiameteraxons.Theybelongtotheintermediateor10nmfilamentprotein/genesuperfamilywhichalsoincludesthekeratins,themajorstructuralproteinsofskin.NeurofilamentsaregenerallyregardedasbeingcomposedofthreemajorproteinsubunitsreferredtoasNF-L,NF-MandNF-H.NF-Listhe"low"or"light"subunit,NF-Misthe"medium"or"middle"subunitandNF-Histhe"heavy"or"high"subunit.ThisnomenclatureisbasedontheapparentsizeoftheseproteinsonSDS-PAGEgels;NF-Listypicallyabout68kDa,NF-Misabout150kDaandNF-Hisabout200kDa.Thesethreeproteins,referredtoastheneurofilamenttriplet,werediscoveredinthe1970sasaresultofstudiesofaxonallytransportedproteins(HoffmanandLasek1975).Afourthprotein,α-internexin,a.k.aNF66,waslaterdiscoveredasaproteinco-purifiingwiththeneurofilamenttripletproteins(PachterandLiem1985).Afifthprotein,peripherin,isfoundinsomeneurofilamentsalongwiththeotherfourproteins,mostlyintheperipheralnervoussystem(Portieretal.1983).Neurofilamentsaremajorproteinsofneuronsandareparticularlyconcentratedinaxons.TheproteinNF-Hhassomeveryunusualproperties.Firstlyitcontains50-60backtobackhexa,heptaoroctapeptiderepeatseachcontainingthesequenceLysine-Serine-Proline(KSP).Theserineresiduesinthesepeptiderepeatsarephosporylatedinvivo,andinaxonalneurofilaments,thesesitesareheavilyphosphorylated.ThephosphorylatedformsofNF-Harealsoquiteresistanttoproteases,suggestingthatthisveryabundantproteinmightbeparticularlyresistanttoproteotyticcleavageuponreleasefromdamagedanddiseasedaxons.Thismeansthatdetectionofthisproteinincerebrospinalfluid(CSF)andbloodwouldprovideinformationaboutthedegreeofaxonalinjurywhichhasoccurred(reviewedinPetzold2005).Asrecentlyshown,thisproteincanbedetectedinquitelargeamountsfollowingexperimentalspinalcordandbraininjuryinrats(Shawetal.2005).Levelsofgreaterthan100ng/mlofpNF-Hweredetectableinbloodfollowingseriousspinalcordinjuryandlowerbutstilleasilydetectablelevelswereseeninbloodofanimalsgivenexperimentalbraininjury.Morerecentstudieshaverevealedconsiderableamountsofthisproteininthebloodofmicetransgenicformutationsofhumancopper/zincsuperoxidedismutase1whichareassociatedwithamyotrophiclateralsclerosis(ALS).ThesemicedevelopanaxonaldegenerationpathologysimilartothatseeninhumanswithALS,andbloodpNF-Hlevelscanbeusedtomonitorthis.Interestingly,pNF-Hisdetectablebeforetheonsetofobviousdiseasesymptoms.OtherstudiesshowthatpNF-Hcanbedetectedintissuehomogenates.OtherexperimentsshowthatpNF-Hcanbedetectedintheplasmaofhumanssufferingfromopticneuritis(Petzoldetal2004)andinelevatedlevelsinthecerebrospinalfluid(CSF)ofindividualssufferingfrombraintumorsandstroke(PetzoldandShaw,unpublished). TestPrincipleWiththeChemiconpNF-Hassaysystem,chickenpolyclonalantibodiesgeneratedagainstpNF-H,arepre-coatedontoa96-wellplateandareusedtocapturepNF-Hfromasample.CapturedpNF-HisdetectedusingpNF-Hspecificrabbitpolyclonalantibodiesandagoatanti-rabbitalkalinephosphataseconjugate.AfteradditionofthesubstratesolutiontheamountofpNF-Hisdetermined.ThestandardcurvedemonstratesadirectrelationshipbetweenOpticalDensity(OD)andpNF-Hconcentration:i.e.,thehighertheODthehigherthepNF-Hconcentrationinthesample. ApplicationTheChemiconpNF-HkitisdesignedtomeasuretheamountofpNF-HintissuehomogenatesandBIOLOGicalfluid(serum,plasma,andserum-free)samplesfrommouseandrat.Thereareenoughreagentsincludedinthiskitforone96-wellimmuno-assayplate.Runningduplicatewellsforsamplesandstandardsisrecommended.
MaterialsRequiredbutNotDelivered	1.Multi-channelorrepeatingPipettes 2.Pipettors&tipscapableofaccuratelymeasuring10-1000μL 3.GraduatedSEROlogicalpipettes 4.96-wellmicroplatereadercapableofmeasuringabsorbanceat405nm 5.Graphpaperformanualplottingofdata 6.1.5mLtubes 7.Mechanicalvortex 8.Two1litercontainers 9.Plateshaker(optional) 10.3NNaOH(optional) 11.Distilledordeionizedwater.

ProductInformation
Components	ImmunoplatePre-coatedwithChickenanti-pNF-HPolyclonalAntibody:(PartNo.2006263).One96-wellimmunoplatepre-coatedwithchickenanti-pNF-Hpolyclonalantibodysealedinafoilpouch. Rabbitanti-pNF-HPolyclonalAntibody:(PartNo.2006919).Onevial(100μL)ofrabbitanti-pNF-Hpolyclonalantibody. Goatanti-Rabbit,AlkalinePhosphataseConjugatedAntibody:(PartNo.2006917).Onevial(10μL)ofgoatanti-rabbitalkalinephosphataseconjugate. BovinepNF-HStandard:(PartNo.2006918).Onevial(5.0ng)ofbovinepNF-Hstandard. 20XTBS:(PartNo.2004913).Onebottle(50mL)of20XTBS. 20%Tween-20?:(PartNo.2004915).Onevial(3mL)of20%Tween-20?. ElisaAssayDiluent:(PartNo.2006921)Onebottle(30mL)ofElisaassaydiluentcontaing0.05%Kathonasapreservative,providedinareadytouseformat. pNPPBuffer:(PartNo.2006261).Onebottle(12mL)ofpNPPbufferforusewith50Xp-NPP,providedinareadytouseformat. 50XpNPP:(PartNo.2006262).Onevial(250μL)of50XpNPP(p-nitrophenylphosphatase)alkalinephosphatasesubstrate. PlateCovers-TwoPlateCovers.
Detectionmethod	Chromogenic

ProductInformation

Components

ImmunoplatePre-coatedwithChickenanti-pNF-HPolyclonalAntibody:(PartNo.2006263).One96-wellimmunoplatepre-coatedwithchickenanti-pNF-Hpolyclonalantibodysealedinafoilpouch.
Rabbitanti-pNF-HPolyclonalAntibody:(PartNo.2006919).Onevial(100μL)ofrabbitanti-pNF-Hpolyclonalantibody.
Goatanti-Rabbit,AlkalinePhosphataseConjugatedAntibody:(PartNo.2006917).Onevial(10μL)ofgoatanti-rabbitalkalinephosphataseconjugate.
BovinepNF-HStandard:(PartNo.2006918).Onevial(5.0ng)ofbovinepNF-Hstandard.
20XTBS:(PartNo.2004913).Onebottle(50mL)of20XTBS.
20%Tween-20?:(PartNo.2004915).Onevial(3mL)of20%Tween-20?.
ElisaAssayDiluent:(PartNo.2006921)Onebottle(30mL)ofElisaassaydiluentcontaing0.05%Kathonasapreservative,providedinareadytouseformat.
pNPPBuffer:(PartNo.2006261).Onebottle(12mL)ofpNPPbufferforusewith50Xp-NPP,providedinareadytouseformat.
50XpNPP:(PartNo.2006262).Onevial(250μL)of50XpNPP(p-nitrophenylphosphatase)alkalinephosphatasesubstrate.
PlateCovers-TwoPlateCovers.

Detectionmethod

Chromogenic

StorageandShippingInformation
StorageConditions	Priortousemaintainthekitcomponentsat2°-8°Cand-20°C,asindicated,untilexpirationdate. 1.WashBuffer-AfterdilutionmaintaintheWashSolutionat2°-8°Cforuptoonemonthoruntilexpirationdate. 2.Tween-20?-Storeatroomtemperatureuponreceiptofkit. 3.ElisaAssayBuffer-Aliquotasrequiredtoavoidrepeatedfreeze-thawcycles.Afterthawing,maintaintheElisaAssayBufferat2°-8°Cforuptoonemonth,oruntilexpirationdate. 4.ImmunoplatePre-coatedwithChickenanti-pNF-HPolyclonalAntibody-Unusedstripwellsmaybereturnedtothefoilpouchandstoredat2°-8°Cforuptoonemonth,oruntilexpirationdate.

StorageandShippingInformation

StorageConditions

Priortousemaintainthekitcomponentsat2°-8°Cand-20°C,asindicated,untilexpirationdate.

1.WashBuffer-AfterdilutionmaintaintheWashSolutionat2°-8°Cforuptoonemonthoruntilexpirationdate.

2.Tween-20?-Storeatroomtemperatureuponreceiptofkit.

3.ElisaAssayBuffer-Aliquotasrequiredtoavoidrepeatedfreeze-thawcycles.Afterthawing,maintaintheElisaAssayBufferat2°-8°Cforuptoonemonth,oruntilexpirationdate.

4.ImmunoplatePre-coatedwithChickenanti-pNF-HPolyclonalAntibody-Unusedstripwellsmaybereturnedtothefoilpouchandstoredat2°-8°Cforuptoonemonth,oruntilexpirationdate.

Applications
Application	ThePhosphorylatedNeurofilamentH(pNF-H)kitisasandwichenzymeimmunoassay(EIA),whichmeasurespNF-H.
KeyApplications	ELISA

BiologicalInformation
SpeciesReactivity	Bovine Human Mouse Pig Rat Feline
AnalytesAvailable	PhosphorylatedNeurofilament(pNF-H)
EntrezGeneNumber	NM_021076.2
GeneSymbol	NEFH NF-H NFH KIAA0845
Modifications	Phosphorylation
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.

PhysicochemicalInformation
Sensitivity	Sensitivity:0.0585ng/mL RangeofDetection:0.0293ng/mLto15ng/mL

Dimensions

MaterialsInformation

MaterialsInformation

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Millipore/NS170 | Phosphorylated Neurofilament (pNF-H) Sandwich ELISA Kit/NS170/1 kit96 wells

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