Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S]
The aim of this study was to develop an enzymatic cholesterol staining method to determine HDL subclasses in a polyacrylamide gradient gel electrophoresis, which further allows staining by protein in the same electrophoresis lane. HDLs from 120 healthy individuals were separated through nondenaturin...
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doaj-7a9b91b4d1b34b86a1de993fba81c2342021-04-28T06:04:16ZengElsevierJournal of Lipid Research0022-22752010-06-0151616101617Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S]Paola Toledo-Ibelles0Cynthia García-Sánchez1Nydia Ávila-Vazzini2Elizabeth Carreón-Torres3Carlos Posadas-Romero4Gilberto Vargas-Alarcón5Oscar Pérez-Méndez6Department of Molecular Biology, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoDepartment of Molecular Biology, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoOutpatients Service, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoDepartment of Molecular Biology, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoDepartment of Endocrinology, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoDepartment of Molecular Biology, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoDepartment of Molecular Biology, Instituto Nacional de Cardiología “Ignacio Chávez”, Mexico City, MexicoThe aim of this study was to develop an enzymatic cholesterol staining method to determine HDL subclasses in a polyacrylamide gradient gel electrophoresis, which further allows staining by protein in the same electrophoresis lane. HDLs from 120 healthy individuals were separated through nondenaturing PAGE. HDLs were stained for cholesterol using an enzymatic semisolid mixture. Once the gels were unstained, they were stained again for proteins with Coomassie blue. The proportions of HDL subclasses were determined by densitometry. HDL subclasses were transformed to concentrations using as reference HDL-cholesterol plasma levels. This method is comparable in linearity and reproducibility to Coomassie blue staining, although it provides quantitative data. As expected, HDL size distribution shifted toward larger particles when determined by cholesterol as compared with protein. With this method, we observed different proportions of HDL subclasses between men and women as compared with Coomassie blue staining. We described a method to determine HDL size distribution by enzymatic cholesterol staining on polyacrylamide gels. The method allows the quantification of the cholesterol plasma concentration of each HDL subclass with the possibility to further stain the protein in the same sample. The combination of HDL staining by cholesterol and protein on electrophoresis gels provides information that may have clinical relevance.http://www.sciencedirect.com/science/article/pii/S0022227520410363high-density lipoprotein subclassesenzymatic stainingoptical densitometry analysisHDL compositioncardiovascular risk |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Paola Toledo-Ibelles Cynthia García-Sánchez Nydia Ávila-Vazzini Elizabeth Carreón-Torres Carlos Posadas-Romero Gilberto Vargas-Alarcón Oscar Pérez-Méndez |
spellingShingle |
Paola Toledo-Ibelles Cynthia García-Sánchez Nydia Ávila-Vazzini Elizabeth Carreón-Torres Carlos Posadas-Romero Gilberto Vargas-Alarcón Oscar Pérez-Méndez Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S] Journal of Lipid Research high-density lipoprotein subclasses enzymatic staining optical densitometry analysis HDL composition cardiovascular risk |
author_facet |
Paola Toledo-Ibelles Cynthia García-Sánchez Nydia Ávila-Vazzini Elizabeth Carreón-Torres Carlos Posadas-Romero Gilberto Vargas-Alarcón Oscar Pérez-Méndez |
author_sort |
Paola Toledo-Ibelles |
title |
Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S] |
title_short |
Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S] |
title_full |
Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S] |
title_fullStr |
Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S] |
title_full_unstemmed |
Enzymatic assessment of cholesterol on electrophoresis gels for estimating HDL size distribution and plasma concentrations of HDL subclasses[S] |
title_sort |
enzymatic assessment of cholesterol on electrophoresis gels for estimating hdl size distribution and plasma concentrations of hdl subclasses[s] |
publisher |
Elsevier |
series |
Journal of Lipid Research |
issn |
0022-2275 |
publishDate |
2010-06-01 |
description |
The aim of this study was to develop an enzymatic cholesterol staining method to determine HDL subclasses in a polyacrylamide gradient gel electrophoresis, which further allows staining by protein in the same electrophoresis lane. HDLs from 120 healthy individuals were separated through nondenaturing PAGE. HDLs were stained for cholesterol using an enzymatic semisolid mixture. Once the gels were unstained, they were stained again for proteins with Coomassie blue. The proportions of HDL subclasses were determined by densitometry. HDL subclasses were transformed to concentrations using as reference HDL-cholesterol plasma levels. This method is comparable in linearity and reproducibility to Coomassie blue staining, although it provides quantitative data. As expected, HDL size distribution shifted toward larger particles when determined by cholesterol as compared with protein. With this method, we observed different proportions of HDL subclasses between men and women as compared with Coomassie blue staining. We described a method to determine HDL size distribution by enzymatic cholesterol staining on polyacrylamide gels. The method allows the quantification of the cholesterol plasma concentration of each HDL subclass with the possibility to further stain the protein in the same sample. The combination of HDL staining by cholesterol and protein on electrophoresis gels provides information that may have clinical relevance. |
topic |
high-density lipoprotein subclasses enzymatic staining optical densitometry analysis HDL composition cardiovascular risk |
url |
http://www.sciencedirect.com/science/article/pii/S0022227520410363 |
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