Peroxiredoxin3,alsoknownasPRDX3,isamemberoftheperoxiredoxinfamilyofantioxidantenzymes,whichreducehydrogenperoxideandalkylhydroperoxides.Peroxiredoxin3isspecificallylocalizedinmitochondriaandbelievedtoplayimportantrolesintheregulationofcellularredoxstatusbyservingasaprimarylineofdefenseagainstH2O2producedduringrespiration.||Recombinantproteincorrespondingtoaa63-256fromhumanPeroxiredoxin3protein,expressedinE.coli||AminoAcidSequence:|MPAVTQHAPYFKGTAVVNGEFKDLSLDDFKGKYLVLFFYPLDFTFVCPTEIVAFSDKANEFHDVNCEVVAVSVDSHFSHLAWINTPRKNGGLGHMNIALLSDLTKQISRDYGVLLEGSGLALRGLFIIDPNGVIKHLSVNDLPVGRSVEETLRLVKAFQYVETHGEVCPANWTPDSPTIK|PSPAASKEYFQKVNQ||SpecifIcActivity:|AsReported.EnzymaticactivitywasconfirmedbymeasuringtheremainingperoxideafterincubationofP3352-22withperoxidefor20minutesatRT.Specificactivityisdefinedastheamountofhydroperoxidethat1ugofenzymecanreduceat25ºCfor1minutes.||ActivityAssay:|1.Preparea50ulreactionmixintoasuitablecontainer:Thefinalconcentrationsare1mMDTT,0.03XPBS,0.5%glycerol.|2.Add5ulofP3352-22withvariousconcentrations(0.25ug,0.5ug)in45ulreactionbuffer.|3.Incubateat25°Cfor2minutes.|4.Add5ulof5mMH2O2asasubstrateandincubatethemixturefor20minutes.|5.Add20ulof26%trichloroaceticacid(TCA)tostopthereaction.|6.Add30ulofFormationsolution(10mMFerrousammoniumsulfate(Fe(II)(NH4)2(SO4)2),2.5MKSCN)|7.RecordtheincreaseinA475nm. ||StorageandStABIlity:|Aliquottoavoidrepeatedfreezingandthawingandstoreat-70°C.Aliquotsarestablefor6monthsafterreceipt.Formaximumrecoveryofproduct,centrifugetheoriginalvialafterthawingandpriortoremovingthecap.

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