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

doi:10.1016/S0021-9673(96)00877-1

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

Division of Laboratory and Genomic Medicine

Research output: Contribution to journal › Article › peer-review

10 Scopus citations

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 language	English
Pages (from-to)	65-70
Number of pages	6
Journal	Journal of Chromatography A
Volume	763
Issue number	1-2
DOIs	https://doi.org/10.1016/S0021-9673(96)00877-1
State	Published - Feb 28 1997

Keywords

HMG14
HMG17
Histone 113
LC
Proteins
Stationary phases

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10.1016/S0021-9673(96)00877-1

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

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.",

keywords = "HMG14, HMG17, Histone 113, LC, Proteins, Stationary phases",

author = "Zhelev, {Nikolai Zh} and Barratt, {Michael J.} and Mahadevan, {Louis C.}",

note = "Funding Information: N. Zh. is fundedb y a BBSRC granta ndM.B. by a Wellcome Trust grant. We are indebted to Dr. Alastair Aitken (NIMR, Mill Hill) for assistance with mass spectrometrayn d to Dr. Alison Clayton (King's College London)f or valuabled iscussionosn testing biological activity of histones and HMG proteins.",

year = "1997",

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doi = "10.1016/S0021-9673(96)00877-1",

language = "English",

volume = "763",

pages = "65--70",

journal = "Journal of Chromatography A",

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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 journal › Article › peer-review

TY - JOUR

T1 - Use of reversed-phase high-performance liquid chromatography on polystyrene-divinylbenzene columns for the rapid separation and purification of acid-soluble nuclear proteins

AU - Zhelev, Nikolai Zh

AU - Barratt, Michael J.

AU - Mahadevan, Louis C.

N1 - Funding Information: N. Zh. is fundedb y a BBSRC granta ndM.B. by a Wellcome Trust grant. We are indebted to Dr. Alastair Aitken (NIMR, Mill Hill) for assistance with mass spectrometrayn d to Dr. Alison Clayton (King's College London)f or valuabled iscussionosn testing biological activity of histones and HMG proteins.

PY - 1997/2/28

Y1 - 1997/2/28

N2 - 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.

AB - 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.

KW - HMG14

KW - HMG17

KW - Histone 113

KW - LC

KW - Proteins

KW - Stationary phases

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U2 - 10.1016/S0021-9673(96)00877-1

DO - 10.1016/S0021-9673(96)00877-1

M3 - Article

C2 - 9129316

AN - SCOPUS:0031588711

SN - 0021-9673

VL - 763

SP - 65

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JO - Journal of Chromatography A

JF - Journal of Chromatography A

IS - 1-2

ER -

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

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