USE OF REVERSED-PHASE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY ON POLYSTYRENE-DIVINYLBENZENE COLUMNS FOR THE RAPID SEPARATION AND PURIFICATION OF ACID-SOLUBLE NUCLEAR PROTEINS

The suitability of polystyrene-divinylbenzene reversed-phase HPLC colu mns for rapid separation and purification of acid-soluble nuclear prot eins was evaluated. We used a polystyrene-divinylbenzene reversed-phas e HPLC column (PLRP-S) for purification of nuclear proteins extracted with 0.3 M HCl or 5% HClO4. We are able to obtain electrophoretically pure fractions for a number of nuclear proteins including HMG14, HMG17 and variants of histone H3. The identity of proteins in these fractio ns was confirmed by immunochemical analysis, protein sequencing, mass spectrometry and migration on two-dimensional polyacrylamide gel elect rophoresis. These methods do not require special preparation of the sa mple and are quicker than similar published methods.