Improvements in two-dimensional gel electrophoresis by utilizing a low cost "in-house" neutral pH sodium dodecyl sulfate-polyacrylamide gel electrophoresis system

被引：28

作者：

Graham, DRM

Garnham, CP

Fu, Q

Robbins, J

Van Eyk, JE

机构：

[1] Johns Hopkins Univ, Sch Med, JHU Proteom Ctr, Dept Pharmacol & Mol Sci, Baltimore, MD 21224 USA

[2] Johns Hopkins Univ, Sch Med, Dept Med, Div Cardiol, Baltimore, MD 21224 USA

[3] Cincinnati Childrens Hosp, Dept Pediat, Div Mol Cardiovasc Biol, Cincinnati, OH USA

来源：

PROTEOMICS | 2005年 / 5卷 / 09期

关键词：

Bis-Tris; casdiac; human immunodeficiency virus type 1; sodium dodecyl sulfate-polyacrylamide gel electrophoresis;

D O I：

10.1002/pmic.200401249

中图分类号：

Q5 [生物化学];

学科分类号：

071010 [生物化学与分子生物学]; 081704 [应用化学];

摘要：

Two-dimensional gel electrophoresis (2-DE) is widely used for initial protein separation in proteomics. Commercial products using neutral PH sodium dodecyl sulfate-polyacrylamide gel electrophoresis ((SDS-PAGE)/(Bis (2-hydroxyethyl) imino-tris (hydroxymethyl) methane-HCl, or Bis-Tris)) have greatly improved this technique, but cost and limited sizes restrict their applications. An "in-house" system is presented, resulting in better resolution, separation, and new spot visualization and improved resolution when compared to Tris-HCl gels. Their utility is demonstrated using albumin-depleted serum samples, rabbit heart left ventricle, and human immunodeficiency virus type 1 (HIV-1).

引用

收藏

页码：2309 / 2314

页数：6

相关论文

共 17 条

[1]

Proteomic analysis of pharmacologically preconditioned cardiomyocytes reveals novel phosphorylation of myosin light chain 1 [J].

Arrell, DK ;

Neverova, I ;

Fraser, H ;

Marbán, E ;

Van Eyk, JE .

CIRCULATION RESEARCH, 2001, 89 (06) :480-487

[2]

Modification of the laemmli sodium dodecyl sulfate-polyacrylamide gel electrophoresis procedure to eliminate artifacts on reducing and nonreducing gels [J].

CannonCarlson, S ;

Tang, J .

ANALYTICAL BIOCHEMISTRY, 1997, 246 (01) :146-148

[3]

Cholesterol depletion of human immunodeficiency virus type 1 and simian immunodeficiency virus with β-cyclodextrin inactivates and permeabilizes the virions:: Evidence for virion-associated lipid rafts [J].

Graham, DRM ;

Chertova, E ;

Hilburn, JM ;

Arthur, LO ;

Hildreth, JEK .

JOURNAL OF VIROLOGY, 2003, 77 (15) :8237-8248

[4]

MULTIPHASIC ZONE ELECTROPHORESIS .1. STEADY-STATE MOVING-BOUNDARY SYSTEMS FORMED BY DIFFERENT ELECTROLYTE COMBINATIONS [J].

JOVIN, TM .

BIOCHEMISTRY, 1973, 12 (05) :871-878

[5]

MULTIPHASIC ZONE ELECTROPHORESIS .2. DESIGN OF INTEGRATED DISCONTINUOUS BUFFER SYSTEMS FOR ANALYTICAL AND PREPARATIVE FRACTIONATION [J].

JOVIN, TM .

BIOCHEMISTRY, 1973, 12 (05) :879-890

[6]

MULTIPHASIC ZONE ELECTROPHORESIS .3. FURTHER ANALYSIS AND NEW FORMS OF DISCONTINUOUS BUFFER SYSTEMS [J].

JOVIN, TM .

BIOCHEMISTRY, 1973, 12 (05) :890-898

[7]

MULTIPHASIC ZONE ELECTROPHORESIS .4. DESIGN AND ANALYSIS OF DISCONTINUOUS BUFFER SYSTEMS WITH A DIGITAL-COMPUTER [J].

JOVIN, TM .

ANNALS OF THE NEW YORK ACADEMY OF SCIENCES, 1973, 209 (JUN15) :477-496

[8]

CLEAVAGE OF STRUCTURAL PROTEINS DURING ASSEMBLY OF HEAD OF BACTERIOPHAGE-T4 [J].

LAEMMLI, UK .

NATURE, 1970, 227 (5259) :680-+

[9]

MONO- AND BIS(2-HYDROXYETHYL)IMINO-TRIS(HYDROXYMETHYL)-METHANE MONO-TRIS AND BIS-TRIS - NEW BUFFER BASES WITH PKAT 7.83 AND 6.46 [J].

LEWIS, JC .

ANALYTICAL BIOCHEMISTRY, 1966, 14 (03) :495-&

[10]

Three isoforms of cyclophilin A associated with human immunodeficiency virus type 1 were found by proteomics by using two-dimensional gel electrophoresis and matrix-assisted laser desorption ionization-time of flight mass spectrometry [J].

Misumi, S ;

Fuchigami, T ;

Takamune, N ;

Takahashi, I ;

Takama, M ;

Shoji, S .

JOURNAL OF VIROLOGY, 2002, 76 (19) :10000-10008