Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting
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AutorRivero Gutiérrez, Belén; Anzola Santander, Andrea; Martínez Augustín, María Olga; Sánchez De Medina López-Huertas, Fermín
Western blotLoading controlPonceau stainingHousekeeping proteinsTGX Stain-Free gels
Rivero-Gutiérrez, B., Anzola, A., Martínez-Augustin, O., & de Medina, F. S. (2014). Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting. Analytical biochemistry, 467, 1-3. [https://doi.org/10.1016/j.ab.2014.08.027]
PatrocinadorSpanish Government SAF2011-22922 SAF2011-22812; Junta de Andalucia CTS6736 CTS164; Instituto de Salud Carlos III
It is currently a routine practice to require a measurement of a housekeeping reference, including actin, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, among others, in Western blots, as it is the rule in RNA blots. Reversible Ponceau staining has been applied successfully to check equal loading of gels. Here we test a new technique, with the Stain-Free gels from Bio-Rad, against both Ponceau staining and housekeeping protein immunodetection under different conditions. Our results show that Stain-Free gels outperform Ponceau staining and that both are more consistent than housekeeping proteins as a loading control.