Moreover, seven of nine model enzymes, including four Zn 2+ proteins that were subjected to NSDS-PAGE retained activity. Retention of Zn 2+ bound in proteomic samples increased from 26 to 98% upon shifting from standard to modified conditions. The modified conditions were called native (N)SDS-PAGE. Reduction of SDS in the running buffer from 0.1% to 0.0375% together with deletion of EDTA also made little impact on the quality of the electrophoretograms of fractions of pig kidney (LLC-PK 1) cell proteome in comparison with that achieved with the SDS-PAGE method. Removal of SDS and EDTA from the sample buffer together with omission of a heating step had no effect on the results of PAGE. To address the need for a high resolution PAGE method that results in the separation of native proteins, experiments tested the impact of changing the conditions of SDS-PAGE on the quality of protein separation and retention of functional properties. This method retains functional properties but at the cost of protein resolving power. To address this shortcoming, blue-native (BN)-PAGE has been introduced. Although covalent structural features of resolved proteins can be determined with SDS-PAGE, functional properties are destroyed, including the presence of non-covalently bound metal ions. The method initially denatures the proteins that will undergo electrophoresis. Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is commonly used to obtain high resolution separation of complex mixtures of proteins.

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