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 columns for rapid separation and purification of acid-soluble nuclear proteins was evaluated. We used a polystyrene-divinylbenzene reversed-phase 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 fractions was confirmed by immunochemical analysis, protein sequencing, mass spectrometry and migration on two-dimensional polyacrylamide gel electrophoresis. These methods do not require special preparation of the sample and are quicker than similar published methods.
Zhelev
N Z
NZ
Nuclear Signalling Laboratory, Randall Institute, King's College London, UK.
Barratt
M J
MJ
Mahadevan
L C
LC
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006609:High Mobility Group Proteins / Q000302:isolation & purification; D006657:Histones / Q000302:isolation & purification; D006851:Hydrochloric Acid; D006863:Hydrogen-Ion Concentration; D009687:Nuclear Proteins / Q000302:isolation & purification*; D011137:Polystyrenes; D012995:Solubility; D014753:Vinyl Compounds
D006609:High Mobility Group Proteins; D006657:Histones; D009687:Nuclear Proteins; D011137:Polystyrenes; D014753:Vinyl Compounds; C004985:divinyl benzene; D006851:Hydrochloric Acid