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Anal Biochem. 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.

Author information

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.
3
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.

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

PMID:
25193447
DOI:
10.1016/j.ab.2014.08.027
[Indexed for MEDLINE]
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