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

Bass, Joseph J. and Wilkinson, Daniel J. and Rankin, Debbie and Phillips, Bethan E. and Szewczyk, Nathaniel J. and 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

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Abstract

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.

Item Type: Article
RIS ID: https://nottingham-repository.worktribe.com/output/796489
Keywords: Westen blot, physiology, SDS PAGE, skeletal muscle
Schools/Departments: University of Nottingham, UK > Faculty of Medicine and Health Sciences > School of Medicine > Division of Medical Sciences and Graduate Entry Medicine
Identification Number: https://doi.org/10.1111/sms.12702
Depositing User: Bass, Joseph
Date Deposited: 13 Jun 2016 13:42
Last Modified: 04 May 2020 17:57
URI: http://eprints.nottingham.ac.uk/id/eprint/33950

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