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Organelles Proteomics


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

Areeba Ahmad
  • Biochemical and Clinical Genetics Laboratory, Section of Genetics, Department of Zoology, Aligarh Muslim University, Aligarh-202 002 (U.P), India
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Riaz Ahmad
  • Corresponding author
  • Biochemical and Clinical Genetics Laboratory, Section of Genetics, Department of Zoology, Aligarh Muslim University, Aligarh-202 002 (U.P), India
  • Email
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2014-04-28 | DOI: https://doi.org/10.2478/orpr-2014-0001

Abstract

Two-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.

Keywords : Urea-SDS-Two dimensional electrophoresis; Liver proteome; Hepatic Fibrosis; Native PAGE; Isoelectric focusing

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About the article

Received: 2014-01-20

Accepted: 2014-04-24

Published Online: 2014-04-28

Published in Print: 2014-01-01


Citation Information: Organelles Proteomics, Volume 1, Issue 1, ISSN (Online) 2084-722X, DOI: https://doi.org/10.2478/orpr-2014-0001.

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© 2014 Areeba Ahmad, Riaz Ahmad . This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License. BY-NC-ND 3.0

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