ABSTRACT
The information obtained from the ground work will pay dividends when decisions have to be made on the choice of resin and starting/elution conditions for chromatography. The initial groundwork will lay the foundations for subsequent experiments; during this process the operator will gain knowledge about a suitable assay and the storage conditions to use with the target protein. Polyphenol oxidase enzymes have a requirement for copper ions so the inclusion of a copper chelating agent such as diethyldithiocarbamate (DIECA) that will help to inhibit their activity. The addition of low concentrations of enzyme substrates, inhibitors, activators and cofactors into the extraction buffer may shift a protein into a compact or stable conformation that may improve the recovery during the extraction or purification process. The process of extracting proteins from cells can sometimes result in a rapid increase in viscosity due to the liberation of nucleic acids into the extraction buffer.
