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Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction

The human placenta is a difficult tissue to work with using proteomic technology since it contains large amounts of lipids and glycogen. Both lipids and glycogen are known to interfere with the first step in the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), the isoelectric focusing....

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Detalles Bibliográficos
Autores principales: Centlow, Magnus, Hansson, Stefan R., Welinder, Charlotte
Formato: Texto
Lenguaje:English
Publicado: Hindawi Publishing Corporation 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2742651/
https://www.ncbi.nlm.nih.gov/pubmed/19756160
http://dx.doi.org/10.1155/2010/458748
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author Centlow, Magnus
Hansson, Stefan R.
Welinder, Charlotte
author_facet Centlow, Magnus
Hansson, Stefan R.
Welinder, Charlotte
author_sort Centlow, Magnus
collection PubMed
description The human placenta is a difficult tissue to work with using proteomic technology since it contains large amounts of lipids and glycogen. Both lipids and glycogen are known to interfere with the first step in the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), the isoelectric focusing. In order to gain the best possible protein separation on 2D-PAGE, an optimized sample preparation protocol for placental proteins was developed. Two different buffers, urea/CHAPS and Hepes, were used for solubilization in combination with six different precipitation methods. The removal of glycogen from the samples by centrifugation was crucial for the final proteome maps. Solubilization with urea/CHAPS in combination with dichloromethane/methanol or acidified acetone proved to be the best precipitation procedures. When applied to clinical placenta samples apolipoprotein A1 was found to be accumulated in the preeclamptic placenta, where it may either have a nutritional effect or act as a modifier of signal transduction.
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spelling pubmed-27426512009-09-15 Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction Centlow, Magnus Hansson, Stefan R. Welinder, Charlotte J Biomed Biotechnol Methodology Report The human placenta is a difficult tissue to work with using proteomic technology since it contains large amounts of lipids and glycogen. Both lipids and glycogen are known to interfere with the first step in the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), the isoelectric focusing. In order to gain the best possible protein separation on 2D-PAGE, an optimized sample preparation protocol for placental proteins was developed. Two different buffers, urea/CHAPS and Hepes, were used for solubilization in combination with six different precipitation methods. The removal of glycogen from the samples by centrifugation was crucial for the final proteome maps. Solubilization with urea/CHAPS in combination with dichloromethane/methanol or acidified acetone proved to be the best precipitation procedures. When applied to clinical placenta samples apolipoprotein A1 was found to be accumulated in the preeclamptic placenta, where it may either have a nutritional effect or act as a modifier of signal transduction. Hindawi Publishing Corporation 2010 2009-09-13 /pmc/articles/PMC2742651/ /pubmed/19756160 http://dx.doi.org/10.1155/2010/458748 Text en Copyright © 2010 Magnus Centlow et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Report
Centlow, Magnus
Hansson, Stefan R.
Welinder, Charlotte
Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction
title Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction
title_full Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction
title_fullStr Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction
title_full_unstemmed Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction
title_short Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction
title_sort differential proteome analysis of the preeclamptic placenta using optimized protein extraction
topic Methodology Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2742651/
https://www.ncbi.nlm.nih.gov/pubmed/19756160
http://dx.doi.org/10.1155/2010/458748
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