Analysis and quantitation of biotinylated apoB-containing lipoproteins with streptavidin-Cy3
| Autor: | Ronald M. Krauss, Michael La Belle, K. Berneis, Patricia J. Blanche |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
Very low-density lipoprotein
Apolipoprotein B gradient gel electrophoresis Lipoproteins QD415-436 Lipoprotein particle Biochemistry LDL chemistry.chemical_compound Endocrinology Rosaniline Dyes apolipoprotein B Humans Biotinylation Apolipoproteins B Fluorescent Dyes Gel electrophoresis Chromatography biology Chemistry Coomassie Brilliant Blue Cell Biology Carbocyanines Molecular biology fluorophor biology.protein Density gradient ultracentrifugation lipids (amino acids peptides and proteins) Streptavidin Azo Compounds Lipoprotein |
| Zdroj: | Journal of Lipid Research, Vol 43, Iss 7, Pp 1155-1159 (2002) |
| ISSN: | 0022-2275 |
| Popis: | Non denaturing gradient gel electrophoresis (GGE) is commonly used to analyze the size distribution of lipoprotein particles. Its relatively low sensitivity and linear dynamic range limit use of GGE to quantify protein content of lipoproteins. We demonstrate a new high sensitivity method for analysis and quantitation of biotinylated apoli- poprotein B (apoB)-containing lipoproteins using a fluores- cent streptavidin-Cy3 conjugate and non covalent preelec- trophoretic binding. Forty-four lipoprotein subfractions spanning the VLDL and LDL particle spectrum subfrac- tions (11 each from four human subjects) were prepared by density gradient ultracentrifugation. An aliquot of each sample was biotinylated and GGE was performed. Gels also were stained for lipid with Oil Red O (32 samples) and for protein with Coomassie Brilliant Blue (30 samples). There was a significant relationship between the Cy3 fluorescent label area under the curve and the mass of apoB ( P � 0.02- 0.004) and total cholesterol ( P � 0.03-0.004). Particle di- ameters of each absorbence/fluorescent peak were com- parable between Oil-Red O and streptavidin-Cy3 treated biotinylated lipoproteins ( � 3.54 A, P � 0.3). Biotinyl- ation and prestaining of lipoprotein particle with streptavi- din-Cy3 provides a new fluorescence-based method for de- tection and quantitative analysis of lipoprotein subspecies by gradient gel electrophoresis. —Berneis, K., M. La Belle, P. J. Blanche, and R. M. Krauss. Analysis and quantitation of biotinylated apoB-containing lipoproteins with Cy3-strepta- vidin. J. Lipid Res. 2002. 43: 1155-1159. |
| Databáze: | OpenAIRE |
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