Description
RecombinantHumanEGFproducedinE.Coliisasingle,non-glycosylated,polypeptidechaincontaining53 aminoacidsandhavingamolecularmassof6222 Dalton.TherHuEGFispurifiedbyproprietarychromatographictechniques.
AminoAcidSequence
ThesequenceofthefirstfiveN-terminalaminoacidswasdeterminedandwasfoundtobeAsn-Ser-Asp-Ser-Glu,whichagreeswiththesequenceofnativehumanEGF.N-terminalmethioninehasbeencompletelyremovedenzymatically.
Source
E.coli
Purity
Greaterthan98.0%asdeterminedby:-(a)AnalysisbyRP-HPLC.-(b)Anion-exchangeFPLC.-©Analysisbyreducingandnon-reducingSDS-PAGESilverStainedgel.Dimersandaggregates:Lessthan1%asdeterminedbysilver-stainedSDS-PAGEgelanalysis.
BIOLOGicalActivity
Biovendor´s recombinanthumanEGFisfullybiologicallyactivewhencomparedtostandards.TheED50,calculatedbythedose-dependantproliferationofmurineBALB/c3T3cells(measuredby3H-thymidineuptake)is<0.1 ng/mlcorrespondingtoaspecificactivityof1×107Units/mg.
Endotoxin
Lessthan0.1 ng/µg(IEU/µg)ofrHuEGF.
Formulation
Theproteinwaslyophilizedfromaconcentrated(1mg/mL)solutionwithnoadditives.
Reconstitution
ItisrecommendedtoreconstitutethelyophilizedrHuEGFinsterile18MΩ-cmH2O notlessthan100µg/ml,whichcanthenbefurtherdilutedtootheraqueoussolutions.
Shipping
Atambienttemperature.Uponreceipt,storetheproductatthetemperaturerecommended below.
Storage/Expiration
LyophilizedrHuEGFalthoughstableatroomtemperaturefor3 weeks,shouldbestoreddesiccatedbelow –18°C.UponreconstitutionrHuEGFshouldbestoredat4°Cbetween2–7daysandforfutureusebelow –18°C.Forlongtermstorageitisrecommendedtoaddacarrierprotein(0.1%HSAorBSA).Pleasepreventfreeze-thawcycles.
PhysicalAppearance
Sterilefilteredwhitelyophilized(freeze-dried)powder.