Choice of native or denaturing puri

Choice of native or denaturing purification conditionsTALON resin retains its protein-binding specificity and yield under a variety of purification conditions. It is stable under both denaturing and native (nondenaturing) conditions. Deciding whether to use native or denaturing purification conditions depends on protein location, solubility, accessibility of the his tag, downstream applications, and preservation of biological activity.Native conditionsPurifying a protein under native conditions is the most efficient way to preserve its biological activity, but requires that the protein be soluble. Advantages include:Eliminating the renaturation step at the end of the purification, saving time, and preventing significant loss of activityRetaining the ability to copurify enzyme subunits, cofactors, and associated proteinsDenaturing conditionsBecause proteins that are overexpressed in prokaryotic systems sometimes form insoluble aggregates called inclusion bodies, you may need to purify proteins under denaturing conditions—using strong denaturants such as 6 M guanidinium or 8 M urea to enhance protein solubility. Advantages include:Complete solubilization of inclusion bodies and his-tagged proteinsImproved binding to the matrix and reduced nonspecific binding, due to full exposure of the his tagHis-tagged proteins purified under denaturing conditions can be used directly in subsequent applications, or may need to be renatured and refolded. Protein renaturation and refolding can be performed prior to elution from the column. However, yields of recombinant proteins will be lower than under native conditions, because urea and guanidinium molecules compete with histidines for binding to metal.Use of reducing agentsPurification with TALON resin may be carried out in the presence of β-mercaptoethanol, but not DTT or DTE, to preserve reduced sulfhydryl (-SH) groups that are important for the biological activity and structure of a given protein. TALON provides higher yields than Ni-NTA in the presence of β-mercaptoethanol.More InformationFeaturesEasy-to-use prepacked 1 ml and 5 ml cartridgesMaximize yield of biologically active proteinSuperior purity & high yieldsApplicationsPurified recombinant his-tagged proteins can be used for:CrystallographyFunctional assaysStructural investigationsOther applicationsAdditional product informationPlease see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.