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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...

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Autores principales: Xiang, Yufang, Zheng, Yuanyuan, Liu, Shaobo, Liu, Gang, Li, Zhi, Dong, Weijie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
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.
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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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