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Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)

There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to...

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Main Authors: Tan, A.A., Azman, S.N., Abdul Rani, N.R., Kua, B.C., Sasidharan, S., Kiew, L.V., Othman, N., Noordin, R., Chen, Y.
Format: Article
Language:English
Published: 2011
Subjects:
Q Science (General)
QR Microbiology
RB Pathology
RZ Other systems of medicine
Online Access:http://eprints.um.edu.my/2643/1/Kiew_LV_2011.docx
http://eprints.um.edu.my/2643/
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spelling my.um.eprints.26432014-12-26T05:22:45Z http://eprints.um.edu.my/2643/ Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE) Tan, A.A. Azman, S.N. Abdul Rani, N.R. Kua, B.C. Sasidharan, S. Kiew, L.V. Othman, N. Noordin, R. Chen, Y. Q Science (General) QR Microbiology RB Pathology RZ Other systems of medicine There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to perform additional sample preparation steps to improve the quality of the final results, yet without selectively losing the proteins. To address this, we developed a general method that is suitable for diverse sample types based on phenolchloroform extraction method (represented by TRI reagent). This method was found to yield good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae, Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line, bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit, PRO-PREP TM Protein Extraction Solution and lysis buffer. The most useful protocol allows the extraction and separation of a wide diversity of protein samples that is reproducible among repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the highest protein yield as well as the greatest number of total proteins spots count for all type of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of different extraction methods used for each type of sample. 2011 Article NonPeerReviewed application/msword en http://eprints.um.edu.my/2643/1/Kiew_LV_2011.docx Tan, A.A. and Azman, S.N. and Abdul Rani, N.R. and Kua, B.C. and Sasidharan, S. and Kiew, L.V. and Othman, N. and Noordin, R. and Chen, Y. (2011) Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE). Tropical Biomedicine, 28 (3). pp. 620-629.
institution Universiti Malaya
building UM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Malaya
content_source UM Research Repository
url_provider http://eprints.um.edu.my/
language English
topic Q Science (General)
QR Microbiology
RB Pathology
RZ Other systems of medicine
spellingShingle Q Science (General)
QR Microbiology
RB Pathology
RZ Other systems of medicine
Tan, A.A.
Azman, S.N.
Abdul Rani, N.R.
Kua, B.C.
Sasidharan, S.
Kiew, L.V.
Othman, N.
Noordin, R.
Chen, Y.
Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)
description There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to perform additional sample preparation steps to improve the quality of the final results, yet without selectively losing the proteins. To address this, we developed a general method that is suitable for diverse sample types based on phenolchloroform extraction method (represented by TRI reagent). This method was found to yield good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae, Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line, bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit, PRO-PREP TM Protein Extraction Solution and lysis buffer. The most useful protocol allows the extraction and separation of a wide diversity of protein samples that is reproducible among repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the highest protein yield as well as the greatest number of total proteins spots count for all type of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of different extraction methods used for each type of sample.
format Article
author Tan, A.A.
Azman, S.N.
Abdul Rani, N.R.
Kua, B.C.
Sasidharan, S.
Kiew, L.V.
Othman, N.
Noordin, R.
Chen, Y.
author_facet Tan, A.A.
Azman, S.N.
Abdul Rani, N.R.
Kua, B.C.
Sasidharan, S.
Kiew, L.V.
Othman, N.
Noordin, R.
Chen, Y.
author_sort Tan, A.A.
title Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)
title_short Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)
title_full Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)
title_fullStr Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)
title_full_unstemmed Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE)
title_sort optimal protein extraction methods from diverse sample types for protein profiling by using two-dimensional electrophoresis (2de)
publishDate 2011
url http://eprints.um.edu.my/2643/1/Kiew_LV_2011.docx
http://eprints.um.edu.my/2643/
_version_ 1643686969836830720
score 13.223943

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