Use of reversed-phase high-performance liquid chromatography on polystyrene-divinylbenzene columns for the rapid separation and purification of acid-soluble nuclear proteins

Nikolai Zh Zhelev, Michael J. Barratt, Louis C. Mahadevan*

*Corresponding author for this work

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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 HMGI4, 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.

Original languageEnglish
Pages (from-to)65-70
Number of pages6
JournalJournal of Chromatography A
Volume763
Issue number1-2
DOIs
Publication statusPublished - 28 Feb 1997
Externally publishedYes

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divinyl benzene
Polystyrenes
Distillation columns
High performance liquid chromatography
Reverse-Phase Chromatography
Nuclear Proteins
Purification
High Pressure Liquid Chromatography
Acids
HMGN2 Protein
Protein Sequence Analysis
Electrophoresis, Gel, Two-Dimensional
Electrophoresis
Histones
Mass spectrometry
Mass Spectrometry
Proteins

Cite this

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abstract = "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 HMGI4, 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.",
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Use of reversed-phase high-performance liquid chromatography on polystyrene-divinylbenzene columns for the rapid separation and purification of acid-soluble nuclear proteins. / Zhelev, Nikolai Zh; Barratt, Michael J.; Mahadevan, Louis C.

In: Journal of Chromatography A, Vol. 763, No. 1-2, 28.02.1997, p. 65-70.

Research output: Contribution to journalArticle

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AU - Zhelev, Nikolai Zh

AU - Barratt, Michael J.

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