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 densitometric analysis is comparable with both techniques. Therefore, routine quantitation of Ponceau staining before antibody probing is validated as an alternative to actin blotting.
Romero-Calvo
Isabel
I
Department of Biochemistry, School of Pharmacy, University of Granada, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Campus de Cartuja s/n, 18071 Granada, Spain.
Ocón
Borja
B
Martínez-Moya
Patricia
P
Suárez
María Dolores
MD
Zarzuelo
Antonio
A
Martínez-Augustin
Olga
O
de Medina
Fermín Sánchez
FS
D000199:Actins / Q000032:analysis*; D000818:Animals; D015153:Blotting, Western / Q000379:methods*; D002460:Cell Line; D003106:Colon / Q000737:chemistry; D006801:Humans; D007668:Kidney / Q000737:chemistry; D008099:Liver / Q000737:chemistry; D051379:Mice; D051381:Rats; D013194:Staining and Labeling / Q000379:methods*
D000199:Actins