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Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis
Aims. Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established. Methods and Material. The method involved acetone precipitation followed by protein ex...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Hindawi Publishing Corporation
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4757711/ https://www.ncbi.nlm.nih.gov/pubmed/26966686 http://dx.doi.org/10.1155/2016/5185317 |
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author | Cheng, Hao-Tsai Hsieh, Sen-Yung Sung, Chang-Mu Pai, Betty Chien-Jung Liu, Nai-Jen Chen, Carl PC |
author_facet | Cheng, Hao-Tsai Hsieh, Sen-Yung Sung, Chang-Mu Pai, Betty Chien-Jung Liu, Nai-Jen Chen, Carl PC |
author_sort | Cheng, Hao-Tsai |
collection | PubMed |
description | Aims. Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established. Methods and Material. The method involved acetone precipitation followed by protein extraction using commercially available 2D Clean-Up kit. The effectiveness was evaluated by 2-dimensional electrophoresis (2DE) profiling quality, including number of protein spots and spot distribution. Results. The total protein of bile fluid in benign biliary disorders was 0.797 ± 0.465 μg/μL. The sample preparation method using acetone precipitation first followed by 2D Clean-Up kit protein extraction resulted in better quality of 2DE gel images in terms of resolution as compared with other sample preparation methods. Using this protocol, we obtained approximately 558 protein spots on the gel images and with better protein spots presentation of haptoglobin, serum albumin, serotransferrin, and transthyretin. Conclusions. Protein samples of bile prepared using acetone precipitation followed by 2D Clean-Up kit exhibited high protein resolution and significant protein profile. This optimized protein preparation protocol can effectively concentrate bile proteins, remove abundant proteins and debris, and yield clear presentation of nonabundant proteins and its isoforms on 2-dimensional electrophoresis gel images. |
format | Online Article Text |
id | pubmed-4757711 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Hindawi Publishing Corporation |
record_format | MEDLINE/PubMed |
spelling | pubmed-47577112016-03-10 Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis Cheng, Hao-Tsai Hsieh, Sen-Yung Sung, Chang-Mu Pai, Betty Chien-Jung Liu, Nai-Jen Chen, Carl PC Biomed Res Int Research Article Aims. Bile is an important body fluid which assists in the digestion of fat and excretion of endogenous and exogenous compounds. In the present study, an improved sample preparation for human bile was established. Methods and Material. The method involved acetone precipitation followed by protein extraction using commercially available 2D Clean-Up kit. The effectiveness was evaluated by 2-dimensional electrophoresis (2DE) profiling quality, including number of protein spots and spot distribution. Results. The total protein of bile fluid in benign biliary disorders was 0.797 ± 0.465 μg/μL. The sample preparation method using acetone precipitation first followed by 2D Clean-Up kit protein extraction resulted in better quality of 2DE gel images in terms of resolution as compared with other sample preparation methods. Using this protocol, we obtained approximately 558 protein spots on the gel images and with better protein spots presentation of haptoglobin, serum albumin, serotransferrin, and transthyretin. Conclusions. Protein samples of bile prepared using acetone precipitation followed by 2D Clean-Up kit exhibited high protein resolution and significant protein profile. This optimized protein preparation protocol can effectively concentrate bile proteins, remove abundant proteins and debris, and yield clear presentation of nonabundant proteins and its isoforms on 2-dimensional electrophoresis gel images. Hindawi Publishing Corporation 2016 2016-02-04 /pmc/articles/PMC4757711/ /pubmed/26966686 http://dx.doi.org/10.1155/2016/5185317 Text en Copyright © 2016 Hao-Tsai Cheng et al. https://creativecommons.org/licenses/by/4.0/ 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 | Research Article Cheng, Hao-Tsai Hsieh, Sen-Yung Sung, Chang-Mu Pai, Betty Chien-Jung Liu, Nai-Jen Chen, Carl PC Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis |
title | Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis |
title_full | Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis |
title_fullStr | Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis |
title_full_unstemmed | Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis |
title_short | Optimizing Human Bile Preparation for Two-Dimensional Gel Electrophoresis |
title_sort | optimizing human bile preparation for two-dimensional gel electrophoresis |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4757711/ https://www.ncbi.nlm.nih.gov/pubmed/26966686 http://dx.doi.org/10.1155/2016/5185317 |
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