Effect of protein application mode and acrylamide concentration on the resolution of protein spots separated by two-dimensional gel electrophoresis

Electrophoresis
H LangenM Fountoulakis

Abstract

Two-dimensional gel electrophoresis separates large numbers of proteins in two steps on the basis of differences in their pIs and molecular masses. The separation is usually performed on immobilized pH gradient strips, followed by gradient polyacrylamide gels separating proteins with molecular masses between 5-200 kDa. For the first-dimensional separation the protein samples are usually applied near one end of the strip. Using total soluble protein extracts of the bacterium Haemophilus influenzae, we found that simultaneous sample application at both the basic and the acidic ends of the strip resulted in detection of more and stronger protein spots in comparison with sample application at one end only. Because many proteins of an organism have similar pI and Mr values, an overlapping of protein spots is frequently observed in the second-dimensional separation. The soluble protein fraction of H. influenzae was further separated on gels of constant acrylamide concentration between 7.5% and 15.0%. We found that for proteins of molecular mass within certain ranges, the gels of homogeneous acrylamide concentration provided more efficient spot separation than the gradient gels. The observed improvements in spot resolution may be usef...Continue Reading

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Citations

Nov 5, 1999·Electrophoresis·K KarlssonM Fountoulakis
Mar 22, 2000·Electrophoresis·M FountoulakisG Lubec
Jun 27, 2001·Electrophoresis·M FountoulakisL Suter
Jul 24, 2001·Electrophoresis·K KrapfenbauerM Fountoulakis
Feb 13, 2002·Electrophoresis·Michael FountoulakisLaura Suter
Jul 5, 2006·Breast Cancer Research and Treatment·Eduardo Lasalvia-PriscoJoshemaria Larrañaga
Nov 25, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jörn Glökler, Philipp Angenendt
Mar 31, 2005·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Michael FountoulakisGert Lubec
Mar 1, 2005·Journal of Molecular and Cellular Cardiology·Michael FountoulakisYassemi Capetanaki
Feb 28, 2003·Analytical Biochemistry·Michael Fountoulakis, Jean François Juranville
Aug 30, 2002·Biochemical and Biophysical Research Communications·Seong Hwan KimGert Lubec
Jul 1, 1998·Journal of Chromatography. a·M FountoulakisB Takács
Jan 23, 1999·Journal of Chromatography. a·M Fountoulakis, H W Lahm
Mar 19, 1999·Journal of Chromatography. a·M FountoulakisB Takács
May 22, 2003·Progress in Neurobiology·Gert LubecMichael Fountoulakis
Nov 15, 2003·Journal of Chemical Neuroanatomy·Andreas PeyrlGert Lubec
Nov 30, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Michael Fountoulakis, Laura Suter
Mar 4, 2006·Journal of Proteome Research·Poonam SarkarGovindarajan T Ramesh
Jun 26, 2001·European Journal of Biochemistry·K KrapfenbauerM Fountoulakis
Aug 31, 2001·Biochemical and Biophysical Research Communications·B LubecM Fountoulakis
Feb 7, 2002·Biochemical and Biophysical Research Communications·Talin GulesserianGert Lubec
Feb 13, 2001·Biochemical and Biophysical Research Communications·B C YooG Lubec
Aug 27, 1998·Electrophoresis·M FountoulakisH Langen
Jun 18, 1998·Electrophoresis·M FountoulakisH Langen
Feb 12, 2005·Electrophoresis·George TsangarisMichael Fountoulakis
Jun 18, 2010·Neuroscience Letters·Xin ZhaoLi-Jun Zhong
Apr 7, 2009·European Journal of Pharmacology·Quan LiXiao-Ping Pu
May 11, 2004·Mass Spectrometry Reviews·Michael Fountoulakis
Dec 30, 2003·International Journal of Cancer. Journal International Du Cancer·Byong Chul YooJae-Gahb Park
Mar 24, 2006·Electrophoresis·Michael Fountoulakis, Sophia Kossida
Nov 9, 2005·Rapid Communications in Mass Spectrometry : RCM·Hu ZhouRong Zeng

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