ArticleThe Journal of physiology2013
Important considerations for protein analyses using antibody based techniques: down-sizing Western blotting up-sizes outcomes.
Article in The Journal of physiology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 64 papers.
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Who cites it
64 citing papers in PubMed, 144 citations in OpenAlex.
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- Intense interval training in healthy older adults increases skeletal muscle [Physiological reports · 2017Trial
- The effect of exercise-intensity on skeletal muscle stress kinase and insulin protein signaling.PloS one · 2017Trial
- Investigation of exercise-mimetic bioactive molecules as modulators of MMP activity and expression in cancer cells.Journal of enzyme inhibition and medicinal chemistry · 2026Article
- Quantitative western blotting adapted for expanded sample capacity with non-linear curve fitting.Journal of plant research · 2026Article
- Integrated effects of altered action potentials and calcium release on skeletal muscle force generation in transgenic Huntington's disease mice.Pflugers Archiv : European journal of physiology · 2026Article
- Exercise attenuates stress-related signaling as sensed by higher phosphorylation of small heat shock proteins in skeletal muscle from older individuals.Journal of sport and health science · 2025Article
- Extraction of Total Protein from Cardiomyocytes and Western Blotting Analysis.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Improving rigor and reproducibility in western blot experiments with the blotRig analysis.Scientific reports · 2024Article
- Sensitive Detection of Histones and γ-H2AX by Immunoblotting: Problems and Solutions.Chemical research in toxicology · 2024Article
- Anti-proliferative and apoptotic effect of cannabinoids on human pancreatic ductal adenocarcinoma xenograft in BALB/c nude mice model.Scientific reports · 2024Article
- Western blotting (immunoblotting): history, theory, uses, protocol and problems.BioTechniques · 2023Review
- An unexpected hydratase synthesizes the green light-absorbing pigment fucoxanthin.The Plant cell · 2023Article
- Calpain-3 Is Not a Sodium Dependent Protease and Simply Requires Calcium for Activation.International journal of molecular sciences · 2023Article
- A Simple, Reproducible Procedure for Chemiluminescent Western Blot Quantification.Bio-protocol · 2023Article
- Long non-coding RNA Tug1 modulates mitochondrial and myogenic responses to exercise in skeletal muscle.BMC biology · 2022Article
- Skeletal myopathy in a rat model of postmenopausal heart failure with preserved ejection fraction.Journal of applied physiology (Bethesda, Md. : 1985) · 2022Article
- Ryanodine receptor leak triggers fiber CaScience advances · 2021Article
- Effects of voluntary wheel running on mitochondrial content and dynamics in rat skeletal muscle.Journal of muscle research and cell motility · 2021Article
4 more citing papers are in PubMed but not listed here.
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Western blotting has been used for protein analyses in a wide range of tissue samples for >30 years. Fundamental to Western blotting success are a number of important considerations, which unfortunately are often overlooked or not appreciated. Firstly, lowly expressed proteins may often be better detected by dramatically reducing the amount of sample loaded. Single cell (fibre) Western blotting demonstrates the ability to detect proteins in small sample sizes, 5-10 μg total mass (1-3 μg total protein). That is an order of magnitude less than often used. Using heterogeneous skeletal muscle as the tissue of representation, the need to undertake Western blotting in sample sizes equivalent to single fibre segments is demonstrated. Secondly, incorrect results can be obtained if samples are fractionated and a proportion of the protein of interest inadvertently discarded during sample preparation. Thirdly, quantitative analyses demand that a calibration curve be used. This is regardless of using a loading control, which must be proven to not change with the intervention and also be appropriately calibrated. Fourthly, antibody specificity must be proven using whole tissue analyses, and for immunofluorescence analyses it is vital that only a single protein is detected. If appropriately undertaken, Western blotting is reliable, quantitative, both in relative and absolute terms, and extremely valuable.
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