Reversible Ponceau staining as a loading control alternative to actin in Western blots

It is becoming standard practice to measure a housekeeping gene, typically actin, in Western blots, as it is the rule in RNA blots. We have applied reversible Ponceau staining to check equal loading of gels and measured actin in parallel under different conditions. Our results show that densitometri...

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Bibliographic Details
Published in:Analytical biochemistry 2010-06, Vol.401 (2), p.318-320
Main Authors: Romero-Calvo, Isabel, Ocón, Borja, Martínez-Moya, Patricia, Suárez, María Dolores, Zarzuelo, Antonio, Martínez-Augustin, Olga, de Medina, Fermín Sánchez
Format: Article
Language:English
Subjects:
Actins - analysis
Animals
Blotting, Western - methods
Cell Line
Colon - chemistry
Humans
Kidney - chemistry
Liver - chemistry
Mice
Rats
Staining and Labeling - methods
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Summary:It is becoming standard practice to measure a housekeeping gene, typically actin, in Western blots, as it is the rule in RNA blots. We have applied reversible Ponceau staining to check equal loading of gels and measured actin in parallel under different conditions. Our results show that densitometric analysis is comparable with both techniques. Therefore, routine quantitation of Ponceau staining before antibody probing is validated as an alternative to actin blotting.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2010.02.036