absoluteantibody/Ab00926-10.0 Anti-Sigma1 [5C6]/Ab00926-10.0/ 200 µg

Description:RecombinantmonoclonalantibodytoSigma1.ManufacturedusingAbAb’sRecombinantPlatformwithvariableregions(i.e.specificity)fromthehybridoma5C6.
SpeciesandIsotype:HumanIgG1,kappa
CloneNumber:5C6
UniProtAccessionNumberofTargetProtein:P04506
PublishedApplication(s):IF;NTRL;IP;FC;crystallisation
PublishedSpeciesReactivity:ReovirusstrainT1L
Immunogen:MurineantibodieswereproducedbyimmunisingCAF1micewithreovirusstrainT3D,withsubsequentimmunisationboostsusingUV-inactivatedstrainT1Lreovirus.ImmunisedmiceproducedantibodiesrecognisingbothT3DandT1L.SplenocyteswereobtainedfromimmunisedmiceandfusedwiththeP3Ag8.653myelomacelllinetogeneratehybridomas.
Specificity:Thestructureof5C6againstSigma1wassolvedcrystallographicallyto3.7angstromresolution(Dietrichetal,2017).The5C6FabregionseachcontacttheupperpartoftheSigma1"head",whichformsabeta-spiralrepeatstructure.TheVHCDRscontacttheantigenmoreextensivelythantheVLCDRs,resultinginadiscontinuousbindinginterface.Reovirusesbindtocell-surfaceglycansascoreceptors,andinparticularT1LreovirusisknowntobindtheglycanGM2tofacilitateinfectivity(Reissetal,2012).The5C6bindingsiteonSigma1partiallyoverlapswithitsGM2bindingsite,blockingviralinfectivitybystericallyinhibitingreceptorbinding.5C6boundSigma1withanaffinityof2nM,shownbySPR.Haemagglutinationassaysof5C6showedactivityagainstreovirusstrainT1L,butnotstrainT3D(Stencel-Baerenwaldetal,2015).
ApplicationNotes:5C6wasshownrecentlytoblockreoviralinfectionofL929mousefibroblastsinadose-dependentmanner,usingimmunofluorescencestaining(Dietrichetal,2017).Fibroblastswereincubatedwithvirus-antibodymixtures,andimmunofluorescencestainingofthecellsenableddetectionofinfectivity.5C6ishighlyspecificfortheT1Lreoviralstrain,asopposedtotheT3Dstrainwhichwasalsotested.5C6wasalsousedtoimmunoprecipitatereoviralparticles.
AntibodyFirstPublishedin:
Virginetal.Monoclonalantibodiestoreovirusrevealstructure/functionrelationshipsbetweencapsidproteinsandgeneticsofsusceptibilitytoantibodyaction.JVirol.1991Dec;65(12):6772-81.Noteonpublication:Describesthegenerationofapanelofanti-reovirus(strainT3DandT1L)mAbs,includingclone5C6,usingmouseimmunisationandhybridomageneration.mAbspecificitieswereprobedusingELISAsandWesternblotting.

ProductForm

Purification:ProteinAaffinitypurified
Suppliedin:PBSwithpreservative(0.02%Proclin300)
StorageRecommendation:Storeat4°Cforupto3months.Forlongertermstoragealiquotintosmallvolumesandstoreat-20°C
Concentration:Seeviallabel.