title: Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting.
abstract: 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.
authors.affiliation: Department of Pharmacology, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), School of Pharmacy, Instituto de Investigación Biosanitaria (ibs.GRANADA), University of Granada, 18071 Granada, Spain.
authors.forename: B
authors.initials: B
authors.lastname: Rivero-Gutiérrez
authors.affiliation: Department of Pharmacology, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), School of Pharmacy, Instituto de Investigación Biosanitaria (ibs.GRANADA), University of Granada, 18071 Granada, Spain.
authors.forename: A
authors.initials: A
authors.lastname: Anzola
authors.affiliation: Department of Biochemistry and Molecular Biology II, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), School of Pharmacy, Instituto de Investigación Biosanitaria (ibs.GRANADA), University of Granada, 18071 Granada, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Martínez-Augustin
authors.affiliation: Department of Pharmacology, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), School of Pharmacy, Instituto de Investigación Biosanitaria (ibs.GRANADA), University of Granada, 18071 Granada, Spain. Electronic address: fsanchez@ugr.es.
authors.forename: F Sánchez
authors.initials: FS
authors.lastname: de Medina
mesh_terms: D000199:Actins / Q000032:analysis*; D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000032:analysis*; D008099:Liver / Q000166:cytology / Q000378:metabolism; D011506:Proteins / Q000032:analysis*; D051381:Rats; D017208:Rats, Wistar; D011924:Rats, Zucker; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D000199:Actins; D011506:Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases
keywords: Housekeeping proteins; Loading control; Ponceau staining; TGX Stain-Free gels; Western blot