Description: RecombinantyeastProteinDisulfideIsomeraseisproducedinE.coliasasingle,non-glycosylated,polypeptidechain,containing503aminoacidsandhavingamolecularmassof62.4kDa.ThePDIhasa6His-tagatN-terminusandispurifiedbyproprietarychromatographictechniques. RecombinantYeastProteinDisulfideIsomeraseisinvolvedindisulphide-bondformationandisomerization,aswellasthereductionofdisulphidebondsinproteins.RecombinantPDIhasbeenfoundtohavemoderateeffects(25-fold)ontherateofoxidativefoldingofproteinsinvitro.
Source: E.coli
RecommendedStorageCondition:-20ºC
PhysicalAppearance: Sterilefilteredliquidformorlyophilizedpowder
Formulation: ThePDIprotein(10mg/ml)solutionwaslyophilizedfromPBSpH7
Solubility: ItisrecommendedtoreconstitutethelyophilizedPDIinsterile18MΩ-cmH2O(notlessthan100µg/ml),whichcanthenbefurtherdilutedtootheraqueoussolutions.
StABIlity: LyophilizedProteinDisulfideIsomerase,althoughstableatroomtemperaturefor3weeks,shouldbestoreddesiccatedbelow-18°C.UponreconstitutionHumanPDIshouldbestoredat4°Cbetween2-7daysandforfutureusebelow-18°C.Forlongtermstorageitisrecommendedtoaddacarrierprotein(0.1%HSAorBSA). Pleaseavoidfreeze-thawcycles.
Purity: Greaterthan99.0%asdeterminedbyRP-HPLC.
ReductaseActivity: 0.001650nm/min-2.Bymeasuringtheturbidityincreaseat650nmduetoinsulinreduction(Holmgren,A.(1979)J.Biol.Chem.254,96279632).Theactivityisexpressedastheratiooftheslopeofalinearpartoftheturbiditycurvetothelagtime(Mart´nez-Galisteo,E.,PADIlla,C.A.,Garcia-Alfonso,C.,Lo´pez-Barea,J.,andBarcena,J.A.(1993)Biochimie(Paris)75,803809).
IsomeraseActivity: 0.5µmolactiveRNaseAmin-1µmolPDI-1.Accordingtothere-activationofreducedanddenaturedRNaseA(Lyles,M.M.andGilbert,H.F.(1991)Biochemistry30,613-619). |