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Comparison of Automated and Traditional Western Blotting Methods

Traditional Western blotting is one of the most used analytical techniques in biological research. However, it can be time-consuming and suffer from a lack of reproducibility. Consequently, devices with different degrees of automation have been developed. These include semi-automated techniques and...

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Autores principales: Sormunen, Aino, Koivulehto, Emma, Alitalo, Kari, Saksela, Kalle, Laham-Karam, Nihay, Ylä-Herttuala, Seppo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10142486/
https://www.ncbi.nlm.nih.gov/pubmed/37104025
http://dx.doi.org/10.3390/mps6020043
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author Sormunen, Aino
Koivulehto, Emma
Alitalo, Kari
Saksela, Kalle
Laham-Karam, Nihay
Ylä-Herttuala, Seppo
author_facet Sormunen, Aino
Koivulehto, Emma
Alitalo, Kari
Saksela, Kalle
Laham-Karam, Nihay
Ylä-Herttuala, Seppo
author_sort Sormunen, Aino
collection PubMed
description Traditional Western blotting is one of the most used analytical techniques in biological research. However, it can be time-consuming and suffer from a lack of reproducibility. Consequently, devices with different degrees of automation have been developed. These include semi-automated techniques and fully automated devices that replicate all stages downstream of the sample preparation, including sample size separation, immunoblotting, imaging, and analysis. We directly compared traditional Western blotting with two different automated systems, iBind™ Flex, which is a semi-automated system designed to perform the immunoblotting, and JESS Simple Western™, a fully automated and capillary-based system performing all steps downstream of sample preparation and loading, including imaging and image analysis. We found that a fully automated system can save time and importantly offer valuable sensitivity. This is particularly beneficial for limited sample amounts. The downside of automation is the cost of devices and reagents. Nevertheless, automation can be a good option to increase output and facilitate sensitive protein analyses.
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spelling pubmed-101424862023-04-29 Comparison of Automated and Traditional Western Blotting Methods Sormunen, Aino Koivulehto, Emma Alitalo, Kari Saksela, Kalle Laham-Karam, Nihay Ylä-Herttuala, Seppo Methods Protoc Brief Report Traditional Western blotting is one of the most used analytical techniques in biological research. However, it can be time-consuming and suffer from a lack of reproducibility. Consequently, devices with different degrees of automation have been developed. These include semi-automated techniques and fully automated devices that replicate all stages downstream of the sample preparation, including sample size separation, immunoblotting, imaging, and analysis. We directly compared traditional Western blotting with two different automated systems, iBind™ Flex, which is a semi-automated system designed to perform the immunoblotting, and JESS Simple Western™, a fully automated and capillary-based system performing all steps downstream of sample preparation and loading, including imaging and image analysis. We found that a fully automated system can save time and importantly offer valuable sensitivity. This is particularly beneficial for limited sample amounts. The downside of automation is the cost of devices and reagents. Nevertheless, automation can be a good option to increase output and facilitate sensitive protein analyses. MDPI 2023-04-20 /pmc/articles/PMC10142486/ /pubmed/37104025 http://dx.doi.org/10.3390/mps6020043 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Brief Report
Sormunen, Aino
Koivulehto, Emma
Alitalo, Kari
Saksela, Kalle
Laham-Karam, Nihay
Ylä-Herttuala, Seppo
Comparison of Automated and Traditional Western Blotting Methods
title Comparison of Automated and Traditional Western Blotting Methods
title_full Comparison of Automated and Traditional Western Blotting Methods
title_fullStr Comparison of Automated and Traditional Western Blotting Methods
title_full_unstemmed Comparison of Automated and Traditional Western Blotting Methods
title_short Comparison of Automated and Traditional Western Blotting Methods
title_sort comparison of automated and traditional western blotting methods
topic Brief Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10142486/
https://www.ncbi.nlm.nih.gov/pubmed/37104025
http://dx.doi.org/10.3390/mps6020043
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