scholarly journals Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction

2010 ◽  
Vol 2010 ◽  
pp. 1-9 ◽  
Author(s):  
Magnus Centlow ◽  
Stefan R. Hansson ◽  
Charlotte Welinder
Keyword(s):  
Signal Transduction ◽  
Gel Electrophoresis ◽  
Protein Separation ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Proteome Analysis ◽  
Apolipoprotein A1 ◽  
Two Dimensional ◽  
2D Page ◽  
Nutritional Effect

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.

Proteome analysis of glycoforms: A review of strategies for the microcharacterisation of glycoproteins separated by two-dimensional polyacrylamide gel electrophoresis

1997 ◽  
Vol 18 (3-4) ◽  
pp. 452-460 ◽  
Author(s):  
Nicolle H. Packer ◽  
Anna Pawlak ◽  
Warren C. Kett ◽  
Andrew A. Gooley ◽  
John W. Redmond ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Proteome Analysis ◽  
Polyacrylamide Gel ◽  
Two Dimensional

A New Method for Proteome Screening with Two-Dimensional Urea-Sds Polyacrylamide Gel Electrophoresis

2014 ◽  
Vol 1 (1) ◽  
Author(s):  
Areeba Ahmad ◽  
Riaz Ahmad
Keyword(s):  
Gel Electrophoresis ◽  
Protein Separation ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cost Effective ◽  
Polyacrylamide Gel ◽  
Present System ◽  
Liver Protein ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis

AbstractTwo-dimensional gel electrophoresis (2DE) separating proteins on the basis of their pI and molecular mass remain the best available technique for protein separation and characterization to date. But due to several limitations, including streak formation in IEF gels, partial solubility of proteins, expensive running conditions and relatively longer time taken, a simple urea-SDS-2D polyacrylamide gel electrophoresis (US2DE) is described here. The system is reasonably sensitive, cost effective with good reproducibility. The method described in this paper employs a chaotropic agent, urea, in the first dimension and sodium dodecyl sulphate (SDS), like conventional system, in the second dimension with an addition of polyacrylamide to screen the liver proteome of healthy and chemically induced fibrotic rats. The system separates the protein on the basis of chargeto- mass ratio and clearly demonstrates differential expression in the liver protein repertoire of healthy and fibrotic rats. Moreover, the present system, like other 2D electrophoretic procedures revealed at least 22 novel spots in the investigated tissues. The technique may be utilized for comprehensive proteome screening of any biological sample and would provide an overview to narrow down the candidate proteins or biomarkers.

Protein extraction from mature rice leaves for two-dimensional gel electrophoresis and its application in proteome analysis

PROTEOMICS ◽  
2004 ◽  
Vol 4 (7) ◽  
pp. 1903-1908 ◽  
Author(s):  
Nazrul Islam ◽  
M. Lonsdale ◽  
N. M. Upadhyaya ◽  
T. J. Higgins ◽  
H. Hirano ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Proteome Analysis ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Rice Leaves

scholarly journals Protein extraction method of Metroxylon sagu leaf for high resolution two dimensional gel electrophoresis and comparative proteomics

2019 ◽  
Author(s):  
Mehvish Nisar ◽  
Hasnain Hussain
Keyword(s):  
Gel Electrophoresis ◽  
Extraction Method ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Two Dimensional ◽  
Two Dimensional Gel Electrophoresis ◽  
Fractionation Method ◽  
Metroxylon Sagu ◽  
Time Required

Abstract Objective This study aimed to determine the best protein extraction method of Metroxylon sagu for the two-dimensional gel electrophoresis and the comparative analysis. Results To perform good proteome research, the most critical step is to establish a method that gives the best quality of extracted total proteins. To develop an optimized protein extraction protocol for two-dimensional polyacrylamide gel electrophoresis (2-DE) analysis of Metroxylon sagu, five protein extraction protocols were compared; polyethene glycol (PEG) fractionation method, SDS/phenol method, TCA/acetone method, combination SDS/phenol and TCA/acetone and imidazole method. The PEG fractionation method was found to give the most reproducible gels with the highest number of spots and highest protein concentration followed by SDS/Phenol method. The lowest number of spots were observed in Imidazole method. The PEG fractionation method provides improved resolution and reproducibility of two-dimensional polyacrylamide gel electrophoresis (2-DE) and reduces the time required to analyze samples. Partitioning rubisco by polyethene glycol (PEG) fractionation provides clearer detection of low-abundance protein. Hence the result from this study propose PEG fractionation as the effective protein extraction method for 2-DE proteomic studies of Metroxylon sagu.

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