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. 2014 Dec 15:467:1-3.
doi: 10.1016/j.ab.2014.08.027. Epub 2014 Sep 1.

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

B Rivero-Gutiérrez  1 , A Anzola  1 , O Martínez-Augustin  2 , F Sánchez de Medina  3
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Free article

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

B Rivero-Gutiérrez et al. Anal Biochem. .
Free article

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.

Keywords: Housekeeping proteins; Loading control; Ponceau staining; TGX Stain-Free gels; Western blot.

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