Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 355, Issue 1-2, Pages 76-79Publisher
ELSEVIER
DOI: 10.1016/j.jim.2010.02.006
Keywords
alpha-tubulin; GAPDH; Hsp90; beta-actin; Gel electrophoresis; Cell confluence
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Funding
- Canadian Institutes of Health Research (CIHR)
- Canadian Breast Cancer Foundation
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- Ontario Centres of Excellence
- Breast Cancer Action Kingston
- Clare Nelson bequest fund
- Canadian Breast Cancer Research Alliance
- Ontario Council on Graduate Studies
- Queen's University
- Ministry of Research and Innovation of the Province of Ontario
- US Army breast cancer program
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Western blotting is a powerful technique to characterize a multitude of cellular proteins. As an internal control, the blots are commonly probed for housekeeping gene products. In this communication, we show that cell confluence significantly affects the levels of two such widely used proteins, alpha-tubulin and Glyceraldehyde-3-Phosphate Dehydrogenase. On the other hand the levels of heat-shock protein-90 and beta-actin remained unchanged at a wide range of cell densities, making these proteins into more reliable loading controls. (C) 2010 Elsevier B.V. All rights reserved.
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