This is for reasons relating to the pursuit of an improved understanding of molecular pathways involved in the regulation of transcription and translation by exercise and nutrition in health, ageing and disease. While such WB approaches are routinely used in many fields of biochemical research, the application of the WB to skeletal muscle and exercise physiology is increasing. 2013) and SUMOylation ( Park-Sarge 2010) to name the main applications). 2012), glycosylation ( Péré-Brissaud et al.

For example, the WB can be used to investigate protein abundance, kinase activity, cellular localization, protein-protein interactions, or monitoring of post-translational modifications (i.e., events of cleavage, phosphorylation ( Nairn et al. The Western blot (WB) has diverse applications for investigating regulatory molecular events underpinning energy metabolism, protein turnover and chronic physiological adaptations. We aim to provide the reader with improved expertise to critically conduct, evaluate and troubleshoot the WB process, to produce reproducible and reliable blots. 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. 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. Despite this popularity, also 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. Indeed, the inclusion of WB data is now considered an essential aspect of many such physiological publications to provide mechanistic insight into regulatory processes. resolving the mechanisms underpinning adaptations to exercise). This is none more so than in relation to skeletal muscle physiology (i.e. VCH-Wiley, Oxford, UK.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. (1997) Electrophoresis in Practice: A Guide to Methods and Applications of DNA and Protein Separations. (1997) The role of IL-4 receptor complex in normal and transformed epithelial cells: structural and functional analysis. (1996) Differential expression of gp200-MR6 molecule in benign hyperplasia and down-regulation in invasive carcinoma of the breast. A., Shousha, S., Melcher, D., and Ritter, M. (1996) The theory and practice of gel electrophoresis of interacting macromolecules. (ed.) (1994) Protein Blotting: A Practical Approach. (1986) Electrophoresis: Theory, Techniques and Biochemical and Clinical Applications. Possible implications for the state of proteins in biological membranes. (1970) Binding of dodecyl sulfate to proteins at high binding ratios. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.

Sds page vs western blot vs elisa