The Open Neuroscience Journal

2008, 2 : 36-40
Published online 2008 December 5. DOI: 10.2174/1874082000802010036
Publisher ID: TONEURJ-2-36

Housekeeping Proteins: Limitations as References During Neuronal Differentiation

Zafira Castaño and Robert M. Kypta
CIC bioGUNE, Cell Biology and Stem Cells Unit, Parque Tecnologico de Bizkaia, Edificio 801A, 48160 Derio, Bizkaia, Spain.

ABSTRACT

The use of “Housekeeping proteins” as loading controls in western blots requires that they do not change their level of expression. However, deviations from this rule have been observed. This study investigated the expression of the commonly used proteins HSP60, βI-tubulin, β-actin and GAPDH by western blot during retinoic acid-induced differentiation of the embryonal carcinoma NTera-2 and the neuroblastoma SH-SY5Y cell lines. β-actin was found to be a poor loading control, increasing significantly during differentiation, whereas HSP60 and GAPDH were the more evenly expressed of those analysed. Our results underline the importance of selecting accurate loading controls in this biological context.

Keywords:

Housekeeping protein, western blotting, NTera-2/clone D1.