Description | ThespectrinfamilyofproteinswereoriginallydiscoveredasmajorcomponentsofthesubmembraneousCytoskeletonofosmoticallylysedredbloodcells(1).Thelysedbloodcellscouldbeseenasclearredbloodcellshapedobjectsinthelightmicroscopeandwerereferredtoasredcell"ghosts".Themajorproteinsoftheseghostsprovedtobeactin,ankyrin,band4.1andseveralotherproteins,includingtwomajorbandsrunningatabout240kDaand260kDaonSDS-PAGEgels.Thispairofbandswasnamed"spectrin"sincetheywerediscoveredintheseredbloodcellghosts(1).Laterworkshowedthatsimilarhighmolecularbandswereseeninmembranepreparationsfromothereukaryoticcelltypes.WorkbyLevineandWillarddescribedapairofabout~240-260kDamolecularweightbandswhichweretransportedattheslowestratealongmammalianaxons(2).Theynamedtheseproteins"fodrin"asantibodystudiesshowedthattheywerelocalizedinthesheathundertheaxonalmembrane,butnotinthecoreoftheaxon(2;fodrosisGreekforsheath).Subsequentlyfodrinwasfoundtobeamemberofthespectrinfamilyofproteins,andthespectrinnomenclatureisnownormallyused(3).Spectrinsformtetramersoftwoalphaandtwobetasubunits,withthealphacorrespondingtothelowermolecularweight~240kDabandandthebetacorrespondingtothe~260kDaorinsomecasemuchlargerband.Mostspectrintetramersareabout0.2micronsor200nmlong,andeachalphaandbetasubunithasacelltypespecificexpressionpattern.Thebasicstructureofeachspectrinsubunitisthespectrinrepeat,whichisasequenceofabout110aminoacidswhichdefinesacompactdomaincontainthreecloselypackedalpha-helices.Eachspectrinsubunitcontainsmultiplecopiesofthisrepeat,with20ineachofthealphasubunits.ThebetaI-IVsubunitseachcontain17spectrinrepeats,whilethebetaVsubunit,alsoknownasbeta-heavyspectrin,contains30oftheserepeats.Thevarioussubunitsalsocontainseveralotherkindsoffunctionaldomain,allowingthespectrintetramertointeractwithavarietyofprotein,ionicandlipidtargets.Thealpha-subunitseachcontainonecalmodulinlikecalciumbindingregionandoneSrc-homology3(SH3)domain,anabundantdomaininvolvedinspecificprotein-proteininteractions.ThebetasubunitsallhaveaN-terminalactinbindingdomainandmayalsohaveoneSH3domainandonepleckstrinhomologydomain,amultifunctionaltypeofbindingdomainwhichinbetaIspectrinatleastbindsthemembranelipidPIP2(5).Spectrinsarebelievedtohaveafunctioningivingmechanicalstrengthtotheplasmamembranesincethetetramersassociatewitheachothertoformadensesubmembraneousgeodesicmeshwork(3).Theyalsobindavarietyofothermembraneproteinsandmembranelipids,andtheproteinstheybindtoarethereforethemselveslocalizedinthemembrane.Diseasesmaybeassociatedwithdefectsinoneorotherofthespectrinsubunits(6).Forexample,someformsofhereditaryspherocytosis,thepresenceofsphericalredbloodcellswhicharepronetolysis,canbetracedtomutationsinsomeofthespectrinsubunits(7).Thealpha-IIsubunitiswidelyexpressedintissuesbut,inthenervoussystem,isfoundpredominantlyinneurons.Theantibodycanthereforebeusedtoidentifyneuronsandfragmentsderivedfromneuronalmembranesincellsintissuecultureandinsectionedmaterial. |
BatchNo. | Seeviallabel |
Unitsize | 100µL |
Antigen | TheantibodywasraisedagainstamixoffiverecombinantconstructscontainingtheentireC-terminalregionofhumanalpha-IIspectrin(aminoacids676-2,400). |
AntibodyType | Polyclonal |
Isotype | IgG |
Accession | Q13813SPTAN1_HUMAN |
Producedin | Rabbit |
Purity | Wholeserum. |
Applications | WesternBlotting(WB)andImmunocytochemistry(IC).SuggesteddilutionforWBis1:5,000-10,000and1:500-1,000forIC.Biosensisrecommendsoptimaldilutions/concentrationsshouldbedeterminedbytheenduser. |
Specificity | Theantibodyreactswitha240kDabandbyWesternblotonmousesciaticnerveextract.Minorbandsbelowmaybeseenandthislikelyrepresentsinvivoproteolyticfragmentsofalpha-IIspectrin.Ithasalsobeenusedsuccessfullyforimmunocytochemistry. |
SpeciesAgainst | Human.Otherspeciesnotyettested. |
Form | Lyophilised. |
Reconstitution | Reconstitutewith100µLsterile-filtered,ultrapurewater.Centrifugebrieflytoremoveanyinsolublematerial. |
Storage | Storelyophilised,unopenedvialat2-8°Corlower.Afterreconstitution,preparealiquotsandstoreat-20°Cto-80°CforahigherstABIlity.Avoidfreeze-thawcycles. |
ExpiryDate | 12monthsafterpurchase(unopenedvial). |
GeneralReferences | 1.MarchesiVT&SteersEJr.Selectivesolubilizationofaproteincomponentoftheredcellmembrane.Science159:203-4(1968). 2.LevineJ&WillardM.Fodrin:axonallytransportedpolypeptidesassociatedwiththeinternalperipheryofmanycells.JCellBiol.90:631-42(1981). 3.BennettV&BainesAJ.Spectrinandankyrin-basedpathways:metazoaninventionsforintegratingcellsintotissues.PhysiolRev.81:1353-92(2001). 4.Djinovic-CarugoK,GautelM,YlänneJ&YoungP.Thespectrinrepeat:astructuralplatformforcytoskeletalproteinassemblies.FEBSLett.513:119-23(2002). 5.Wang,DSandShawG.TheassociationoftheC-terminalregionofbetaIsigmaIIspectrintobrainmembranesismediatedbyaPHdomain,doesnotrequiremembraneproteins,andcoincideswithainositol-1,4,5triphosphatebindingsite.BBRC217:608-15(1995). 6.BennettV&HealyJ.Organizingthefluidmembranebilayer:diseaseslinkedtospectrinandankyrin.TrendsMolMed14:28-36(2008).7.EberS&LuxSE.Hereditaryspherocytosis--defectsinproteinsthatconnectthemembraneskeletontothelipidbilayer.SeminHematol41:118-41(2004). |
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