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Millipore/MAB5278 | Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8/MAB52

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貨號: MAB5278

品牌: Millipore

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Description
CatalogueNumber	MAB5278
BrandFamily	Chemicon®
TradeName	Chemicon
Description	Anti-TryptophanHydroxylase/TyrosineHydroxylase/PhenylalanineHydroxylaseAntibody,clonePH8
BackgroundInformation	PH8AntibodyisamurinemonoclonalantibodythatbindsacommonepitopeofTryptophanhydroxylase(TRH),Tyrosinehydroxylase(TYH)andPhenylalaninehydroxylase(PAH).1IntheliteraturethismonoclonalantibodyiscitedasPH8.Tyrosinehydroxylase(TYH)istheenzymewhichconvertstyrosinetodihydroxyphenylalanine(L-dopa),aprecursorofthecatecholamineneurotransmittersdopamine,noradrenalineandadrenaline.Tryptophanhydroxylase(TRH)istheenzymethatconverts5-hydroxytryptophantoSEROtonin.InfreshtissuethePH8AntibodybindstoTYHandTRHsoisaMarkerforTYH-containingcatecholaminergicneurons,2andserotonergicneurons.3,4,5Tryptophanhydroxylase(TRH)canbeusedasamarkerforserotoninasitconverts5-hydroxy-tryptophantoserotonin.Serotoninisrapidlymetabolisedandisunabletobedetectedbyanti-serotoninantibodiesinpostmortemtissue.Inhumantissuethathasbeenformalin-fixed,duetoachangeintheantigenicdeterminantoftyrosinehydroxylase(TYH),PH8Antibodywillbindonlytryptophanhydroxylase(TRH).3,4,5ThePH8Antibodycanthereforebeusedtospecificallyidentifyserotonergicneuronsinfixedhumantissue.Phenylalaninehydroxylase(PAH)canbedetectedinhepatictissuesectionsusingthePH8Antibody.

ProductInformation
Format	Purified
Presentation	TheimmunoglobulinfractionhasbeenpurifiedfromascitesfluidbyProteinGchromatography.500μgliquidproteinat2mg/mLinphosphatebufferedsaline(PBS).Theproductissuppliedsterile-filteredthrougha0.22micronfilter.

StorageandShippingInformation
StorageConditions	ThePH8Antibodyisshippedliquidatambienttemperature.Liquidmaterialisstableforatupto6monthswhenstoredat2-8°C.

Applications
Application	ThisAnti-TryptophanHydroxylase/TyrosineHydroxylase/PhenylalanineHydroxylaseAntibody,clonePH8isvalidatedforuseinIHC,IP,WBforthedetectionofTryptophanHydroxylase/TyrosineHydroxylase/PhenylalanineHydroxylase.
KeyApplications	Immunohistochemistry Immunoprecipitation WesternBlotting
ApplicationNotes	ThePH8Antibodycanbeusedtoidentifydopaminergicandserotonergicneuronsbyimmuno-histochemistryandforWesternblotanalysis,immunoprecipitationandimmuno-histochemistryofTYH,PAHandTRH. PROTOCOLS Immunohistochemistry ThePH8Antibodycanbeusedfortheimmunohistochemicaldetectionofdopaminergicandserotonergicneuronsinhumanandratbrainstemtissue. 1.Tissueshouldbeformalinfixedandstoredinformalinpriortouseforaminimumoffivedays. 2.Cryoprotecttissueusing30%sucrosein0.1MTrispH7.4bufferfor24-72hours. 3.Cutusingasledgemicrotometo50μmthickness. 4.WashthetissuesamplesusingTrisbufferpriortocommencingthestainingprocedure. 5.Treatthetissuesamplesfor3x15minutesin50%alcohol. 6.Treatthetissuesamplesfor20minutesin50%alcoholand3%H2O2. 7.Treattissuefor20minuteswith10%normalhorseseruminTrisbuffer.ThisactstoblockendogeneousH2O2staining. 8.DilutethePH8AntibodyinTrisbuffer,addtothetissuesamplesandincubatefor1-3days.Recommendeddilutionsare1:2,000-1:10,000.Athighantibodyconcentrationallmonoaminergicneuronsarestainedwithnodistinctionbetweenserotonegicandcatecholominergiccells.Bydilutingtheantibodyconcentrate,celltypesaredistinguishableduetovariationinstainingintensity.Atlowerconcentrations(1:5,000-1:10,000dilution)onlyserotonergiccellswillstain. 9.Washtissue(3x15minutes),thenaddabiotinylatedanti-mousesecondaryantibodyandincubateonanorbitalshakeratroomtemperature(RT)for1hour. 10.Washtissue(3x15minutes)andincubateonanorbitalshakeratRTfor1hourwiththetertiarycomplex(ELITEKIT,Vector,USA). 11.Washtissue(3x15minutes)andincubateonanorbitalshakeratRTfor10minuteswithTrisbuffereddiamino-benzidinesubstrate.Add0.1%H2O2andincubateonanorbitalshakeratRTforafurther5minutes. 12.Mounttissueontogelatinisedslidesandallowtodrypriortomicroscopy.

BIOLOGicalInformation
Clone	PH8
Host	Mouse
Isotype	IgG1
SpeciesReactivity	Human
AntibodyType	MonoclonalAntibody
EntrezGeneNumber	NM_004179.1
EntrezGeneSummary	Thisgeneencodesamemberofthepterin-dependentaromaticacidhydroxylasefamily.Theencodedproteincatalyzesthefirstandratelimitingstepinthebiosynthesisofserotonin,animportanthormoneandneurotransmitter.Thehumangenomecontainstworelatedtryptophanhydroxylases,oneonchromosome11p15-p14andoneonchromosome12q21.Thisgeneisexpressedpredominantlyinthegut,pinealgland,spleen,andthymus.
GeneSymbol	TPH1 TPRH MGC119994 TPH TRPH EC1.14.16.4
UniProtNumber	P17752 P07101
UniProtSummary	FUNCTION:SwissProt:P07101#Playsanimportantroleinthephysiologyofadrenergicneurons. COFACTOR:Fe(2+)ion. SIZE:528aminoacids;58524Da TISSUESPECIFICITY:Mainlyexpressedinthebrainandadrenalglands. DISEASE:SwissProt:P07101#DefectsinTHarethecauseofautosomalrecessiveSegawasyndrome[MIM:605407];alsoknownasDOPA-responsivedystonia.Typically,itbeginsinchildhoodoradolescencewithprogressivedifficultyinwalkingand,insomecases,spasticity.Somecasespresentwithparkinsoniansymptomsininfancyandarereferredtoasautosomalrecessiveinfantileparkinsonism. SIMILARITY:SwissProt:P07101##Belongstothebiopterin-dependentaromaticaminoacidhydroxylasefamily.

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Millipore/MAB5278 | Anti-Tryptophan Hydroxylase/Tyrosine Hydroxylase/Phenylalanine Hydroxylase Antibody, clone PH8/MAB52

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