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

Anal Biochem. 2010 Jun 15;401(2):318-20. doi: 10.1016/j.ab.2010.02.036. Epub 2010 Mar 3.


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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Actins / analysis*
  • Animals
  • Blotting, Western / methods*
  • Cell Line
  • Colon / chemistry
  • Humans
  • Kidney / chemistry
  • Liver / chemistry
  • Mice
  • Rats
  • Staining and Labeling / methods*


  • Actins