PolyclonalAntibodytoHumanTopoiosomeraseI(SclerodermaPatientSerum)Description
SomesclerodermapatientsproducehightiterantibodydirectedagainsthumantopoisomeraseI(Sheroetal.,1986). Theimmunoglobulinfractionwaspurifiedfromthesepatientsandshowntoreactspecificallywiththe100kDaformofTopoIusingWesternblotting. Theimmunecomplexesproducedwiththisreagentwillbind[125I]-proteinA. WhenintacttopoisomeraseIisused,asinglepolypeptideof100kDawasdetected;however,sub-bandsproducedbyproteolysiswillalsobindthisreagent,asexpected. TheantibodyisparticularlyusefulasaWesternblottingprobeandcanbeusedathighdilutions(>1:5,000). Crossreactivityofthisantibodywithotherspecieshasbeendemonstrated(withcalfthymus). Importantnoteaboutproductuse:HumantopoisomeraseIundergoesspontaneousdegradationespeciallywithrepeatfreezing/thawing.Theactivityoftheenzymeisnotaffectedsignificantly;however,sub-bandsmaybedetectableinWesternblottingexperiments.Thisantibodymaybeusedinimmunoprecipitationexperiments.

UnitDefinition:
ThePolyclonalAntibodytoHumanTopoiosomeraseIconcentrationisinWesternblottingunits;oneunitcorrespondstoa1:1000dilutionofprobeforaroutineWesternblot(1ml). Forexample,10unitsofantibodywillmake10mlofdilutedprobe(tobeaddeddirectlytoasingleblot).

Reference:
Shero,J.H.Bordwell,B.,Rothfield,N.F.andEarnshaw,W.C.1986.HightitersofautoantibodiestotopoisomeraseI(Scl-70)inserafromsclerodermapatients.Science231:737-740

MaterialSafetyData:
THISPRODUCEISNOTLICENSEDORAPPROVEDFORADMINISTRATIONTOHUMANSORTOANIMALSOTHERTHANEXPERIMENTALANIMALS.

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