Type:Protein
SpeciesReactivity:H
B=Bovine;Ca=Cat;Ch=Chicken;D=Dog;EQ=Equine;GP=GuineaPig;H=Human;M=Mouse;P=Porcine;Pr=Primate;R=Rat;Rb=Rabbit;Y=Yeast;Xe=Xenopus;Ze=Zebrafish;;;;NA-NotApplicable;STP=Step-TactinProteins;All
ACDNAencodingfulllengthhumanUCHL1wasinsertedintoaeukaryoticexpressionvectorwithaC-terminalHis-Tagandsubsequentlytransformedinto E.coli BL21cells.CellsweregrowninLBmediumsupplementedwithampicillinat37°C.Cellswereharvestedafterinducedproteinexpressionfor6hrs.Solubleproteinwaspurifiedusingimmobilizedmetalaffinitychromatography.YieldsofsolubleUCHL1proteinwereestimatedbyperformingSDS-PAGEandstainingwithCoomassieBlueR250.PurifiedproteinwasconcentratedfollowingextensivedialysisinPBSto1mg/mL,andstoredat-70°C(recommendedforlongtermstorage).Forthesubsequentanalyses,theconcentrationofUCHL1wasobtainedusingtheBradfordmethodandconfirmedwithadensitometry-basedImageJquantification.
Image:His-tagged-UCHL1,wasexpressedandpurifiedfromE.coliBL21usingimmobilizedmetalaffinitychromatography.ProteinconcentrationwasdeterminedusingBSAProteinBradford,andconfirmedwithadensitometry-basedImagJquantificationusingBSAasstandard.PictureshowstheCoomassiegelofUCHL1andBSAatserialdilution.UCHL1runsat~37kDa.
