Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting |
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| Authors: | B Rivero-Gutiérrez A Anzola O Martínez-Augustin F Sánchez de Medina |
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| Institution: | 1. 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;2. 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 |
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| 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. |
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| Keywords: | Western blot Loading control Ponceau staining Housekeeping proteins TGX Stain-Free gels |
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