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Differential proteome analysis of the preeclamptic placenta using optimized protein extraction.

Centlow, Magnus LU ; Hansson, Stefan LU and Welinder, Charlotte LU (2010) In Journal of Biomedicine and Biotechnology 2010(Sep 13).
Abstract
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... (More)
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. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biomedicine and Biotechnology
volume
2010
issue
Sep 13
publisher
Hindawi Publishing Corporation
external identifiers
  • wos:000271075100001
  • pmid:19756160
  • scopus:70449715729
ISSN
1110-7251
DOI
10.1155/2010/458748
language
English
LU publication?
yes
id
29d67e30-a75c-4fa2-ac36-101a1b7e6068 (old id 1483439)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/19756160?dopt=Abstract
date added to LUP
2009-10-07 12:02:53
date last changed
2018-06-17 04:54:43
@article{29d67e30-a75c-4fa2-ac36-101a1b7e6068,
  abstract     = {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.},
  articleno    = {458748},
  author       = {Centlow, Magnus and Hansson, Stefan and Welinder, Charlotte},
  issn         = {1110-7251},
  language     = {eng},
  number       = {Sep 13},
  publisher    = {Hindawi Publishing Corporation},
  series       = {Journal of Biomedicine and Biotechnology},
  title        = {Differential proteome analysis of the preeclamptic placenta using optimized protein extraction.},
  url          = {http://dx.doi.org/10.1155/2010/458748},
  volume       = {2010},
  year         = {2010},
}