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Updated: May 10, 2026

V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting
Published on: December 30, 2013
Stain-Free total protein staining is a superior loading control to β-actin for Western blots
Jennifer E Gilda1, Aldrin V Gomes
1Department of Neurobiology, Physiology, and Behavior, University of California, Davis, CA 95616, USA.
Abstract:
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.

