Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting Rivero Gutiérrez, Belén Anzola Santander, Andrea Martínez Augustín, María Olga Sánchez De Medina López-Huertas, Fermín Western blot Loading control Ponceau staining Housekeeping proteins TGX Stain-Free gels The authors want to thank the input of Arnaud Remy and Enrique Orozco of Bio-Rad Laboratories. This study was supported by the Ministry of Science and Innovation (SAF2011-22922 and SAF2011-22812), and by funds from Junta de Andalucia (CTS6736 and CTS164). B. Rivero and A. Anzola were supported by fellowships of the Ministry of Education and Science of Spain and Junta de Andalucia, respectively. CIBERehd is funded by the 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. 2020-07-29T11:50:22Z 2020-07-29T11:50:22Z 2014-12-15 info:eu-repo/semantics/article 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] http://hdl.handle.net/10481/63191 10.1016/j.ab.2014.08.027 eng http://creativecommons.org/licenses/by/3.0/es/ info:eu-repo/semantics/openAccess Atribución 3.0 España Elsevier