An overview of technical considerations for Western blotting applications to physiological research

The applications of Western/immuno-blotting (WB) techniques have reached multiple layers of the scientific community and are now considered routine procedures in the field of physiology. This is none more so than in relation to skeletal muscle physiology (i.e. resolving the mechanisms underpinning a...

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Main Authors: Bass, Joseph J., Wilkinson, Daniel J., Rankin, Debbie, Phillips, Bethan E., Szewczyk, Nathaniel J., Smith, Kenneth, Atherton, Philip J.
Format: Article
Published: Wiley 2016
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Online Access:https://eprints.nottingham.ac.uk/33950/
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author Bass, Joseph J.
Wilkinson, Daniel J.
Rankin, Debbie
Phillips, Bethan E.
Szewczyk, Nathaniel J.
Smith, Kenneth
Atherton, Philip J.
author_facet Bass, Joseph J.
Wilkinson, Daniel J.
Rankin, Debbie
Phillips, Bethan E.
Szewczyk, Nathaniel J.
Smith, Kenneth
Atherton, Philip J.
author_sort Bass, Joseph J.
building Nottingham Research Data Repository
collection Online Access
description The applications of Western/immuno-blotting (WB) techniques have reached multiple layers of the scientific community and are now considered routine procedures in the field of physiology. This is none more so than in relation to skeletal muscle physiology (i.e. resolving the mechanisms underpinning adaptations to exercise). Indeed, the inclusion of WB data is now considered an essential aspect of many such physiological publications to provide mechanistic insight into regulatory processes. Despite this popularity, and due to the ubiquitous and relatively inexpensive availability of WB equipment, the quality of WB in publications and subsequent analysis and interpretation of the data can be variable, perhaps resulting in spurious conclusions. This may be due to poor laboratory technique and/or lack of comprehension of the critical steps involved in WB and what quality control procedures should be in place to ensure robust data generation. The present review aims to provide a detailed description and critique of WB procedures and technicalities, from sample collection through preparation, blotting and detection to analysis of the data collected. We aim to provide the reader with improved expertise to critically conduct, evaluate and troubleshoot the WB process, to produce reproducible and reliable blots.
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spelling nottingham-339502020-05-04T17:57:38Z https://eprints.nottingham.ac.uk/33950/ An overview of technical considerations for Western blotting applications to physiological research Bass, Joseph J. Wilkinson, Daniel J. Rankin, Debbie Phillips, Bethan E. Szewczyk, Nathaniel J. Smith, Kenneth Atherton, Philip J. The applications of Western/immuno-blotting (WB) techniques have reached multiple layers of the scientific community and are now considered routine procedures in the field of physiology. This is none more so than in relation to skeletal muscle physiology (i.e. resolving the mechanisms underpinning adaptations to exercise). Indeed, the inclusion of WB data is now considered an essential aspect of many such physiological publications to provide mechanistic insight into regulatory processes. Despite this popularity, and due to the ubiquitous and relatively inexpensive availability of WB equipment, the quality of WB in publications and subsequent analysis and interpretation of the data can be variable, perhaps resulting in spurious conclusions. This may be due to poor laboratory technique and/or lack of comprehension of the critical steps involved in WB and what quality control procedures should be in place to ensure robust data generation. The present review aims to provide a detailed description and critique of WB procedures and technicalities, from sample collection through preparation, blotting and detection to analysis of the data collected. We aim to provide the reader with improved expertise to critically conduct, evaluate and troubleshoot the WB process, to produce reproducible and reliable blots. Wiley 2016-06-05 Article PeerReviewed Bass, Joseph J., Wilkinson, Daniel J., Rankin, Debbie, Phillips, Bethan E., Szewczyk, Nathaniel J., Smith, Kenneth and Atherton, Philip J. (2016) An overview of technical considerations for Western blotting applications to physiological research. Scandinavian Journal of Medicine & Science in Sports, 27 (1). pp. 4-25. ISSN 1600-0838 Westen blot physiology SDS PAGE skeletal muscle http://onlinelibrary.wiley.com/doi/10.1111/sms.12702/abstract doi:10.1111/sms.12702 doi:10.1111/sms.12702
spellingShingle Westen blot
physiology
SDS PAGE
skeletal muscle
Bass, Joseph J.
Wilkinson, Daniel J.
Rankin, Debbie
Phillips, Bethan E.
Szewczyk, Nathaniel J.
Smith, Kenneth
Atherton, Philip J.
An overview of technical considerations for Western blotting applications to physiological research
title An overview of technical considerations for Western blotting applications to physiological research
title_full An overview of technical considerations for Western blotting applications to physiological research
title_fullStr An overview of technical considerations for Western blotting applications to physiological research
title_full_unstemmed An overview of technical considerations for Western blotting applications to physiological research
title_short An overview of technical considerations for Western blotting applications to physiological research
title_sort overview of technical considerations for western blotting applications to physiological research
topic Westen blot
physiology
SDS PAGE
skeletal muscle
url https://eprints.nottingham.ac.uk/33950/
https://eprints.nottingham.ac.uk/33950/
https://eprints.nottingham.ac.uk/33950/