Housekeeping Genes; Expression Levels May Change with Density of Cultured Cells

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.