Stain-Free total protein staining is a superior loading control to β-actin for Western blots

Anal Biochem. 2013 Sep 15;440(2):186-8. doi: 10.1016/j.ab.2013.05.027. Epub 2013 Jun 6.


Semi-quantification of proteins using Western blots typically involves normalization against housekeeping genes such as β-actin. More recently, Ponceau S and Coomassie blue staining have both been shown to be suitable alternatives to housekeeping genes as loading controls. Stain-Free total protein staining offers the advantage of no staining or destaining steps. Evaluation of the use of Stain-Free staining as an alternative to β-actin or the protein stain Ponceau S showed that Stain-Free staining was superior to β-actin and as good as or better than Ponceau S staining as a loading control for Western blots.

Keywords: Nitrocellulose; Ponceau S; Stain Free; Western blotting; β-Actin.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actins / metabolism*
  • Animals
  • Blotting, Western / methods*
  • Liver / cytology
  • Rats
  • Staining and Labeling / methods*


  • Actins