[HTML][HTML] Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting

B Rivero-Gutiérrez, A Anzola, O Martínez-Augustin… - Analytical …, 2014 - Elsevier
Analytical biochemistry, 2014Elsevier
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 …
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.
Elsevier