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Comparison of the sensitivity of Western blotting between PVDF and NC membranes
Western blotting (WB) is one of the most widely used techniques to identify proteins as well as post translational modifications of proteins. The selection of electroblotted membrane is one of the key factors affecting the detection sensitivity of the protein which is transferred from gel to membran...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8187435/ https://www.ncbi.nlm.nih.gov/pubmed/34103620 http://dx.doi.org/10.1038/s41598-021-91521-8 |
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author | Xiang, Yufang Zheng, Yuanyuan Liu, Shaobo Liu, Gang Li, Zhi Dong, Weijie |
author_facet | Xiang, Yufang Zheng, Yuanyuan Liu, Shaobo Liu, Gang Li, Zhi Dong, Weijie |
author_sort | Xiang, Yufang |
collection | PubMed |
description | Western blotting (WB) is one of the most widely used techniques to identify proteins as well as post translational modifications of proteins. The selection of electroblotted membrane is one of the key factors affecting the detection sensitivity of the protein which is transferred from gel to membrane in WB. The most common used membranes are polyvinylidene fluoride (PVDF) and nitrocellulose (NC) membranes. Which membrane of these two is more suitable for WB has not been reported so far. Here, by incubating proteins which were transferred to PVDF or NC membranes with a series of antibodies and different types of lectins, we investigated the relationship between the binding ability of these two membranes to proteins or glycoproteins and the molecular weight of the target protein. The antibody re-probed ability of the two membranes was also explored. Moreover, we verified the above results by directly incubating proteins having different molecular weights onto PVDF or NC membranes. Bound proteins were stained with direct blue-71, and the staining intensity was quantitated by scanning and densitometry. |
format | Online Article Text |
id | pubmed-8187435 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-81874352021-06-09 Comparison of the sensitivity of Western blotting between PVDF and NC membranes Xiang, Yufang Zheng, Yuanyuan Liu, Shaobo Liu, Gang Li, Zhi Dong, Weijie Sci Rep Article Western blotting (WB) is one of the most widely used techniques to identify proteins as well as post translational modifications of proteins. The selection of electroblotted membrane is one of the key factors affecting the detection sensitivity of the protein which is transferred from gel to membrane in WB. The most common used membranes are polyvinylidene fluoride (PVDF) and nitrocellulose (NC) membranes. Which membrane of these two is more suitable for WB has not been reported so far. Here, by incubating proteins which were transferred to PVDF or NC membranes with a series of antibodies and different types of lectins, we investigated the relationship between the binding ability of these two membranes to proteins or glycoproteins and the molecular weight of the target protein. The antibody re-probed ability of the two membranes was also explored. Moreover, we verified the above results by directly incubating proteins having different molecular weights onto PVDF or NC membranes. Bound proteins were stained with direct blue-71, and the staining intensity was quantitated by scanning and densitometry. Nature Publishing Group UK 2021-06-08 /pmc/articles/PMC8187435/ /pubmed/34103620 http://dx.doi.org/10.1038/s41598-021-91521-8 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Xiang, Yufang Zheng, Yuanyuan Liu, Shaobo Liu, Gang Li, Zhi Dong, Weijie Comparison of the sensitivity of Western blotting between PVDF and NC membranes |
title | Comparison of the sensitivity of Western blotting between PVDF and NC membranes |
title_full | Comparison of the sensitivity of Western blotting between PVDF and NC membranes |
title_fullStr | Comparison of the sensitivity of Western blotting between PVDF and NC membranes |
title_full_unstemmed | Comparison of the sensitivity of Western blotting between PVDF and NC membranes |
title_short | Comparison of the sensitivity of Western blotting between PVDF and NC membranes |
title_sort | comparison of the sensitivity of western blotting between pvdf and nc membranes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8187435/ https://www.ncbi.nlm.nih.gov/pubmed/34103620 http://dx.doi.org/10.1038/s41598-021-91521-8 |
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