title: Western blotting using in-gel protein labeling as a normalization control: stain-free technology.
abstract: Western blotting is a commonly used laboratory technique for semi-quantifying protein amounts. It is important when quantifying protein expression to account for differences in the amount of total protein loaded onto the gel using a loading control. Common loading controls include housekeeping proteins, such as β-actin or GAPDH, quantified by Western blot, or total protein, quantified using a stain such as Coomassie Brilliant Blue or Ponceau S. A more recently developed method for total protein quantification utilizes stain-free technology, which has a linear dynamic detection range and allows for protein detection on both gels and membranes. Here, we describe the theory and use of stain-free gels for total protein quantification and normalization of Western blots.
authors.affiliation: Department of Neurobiology, Physiology, and Behavior, University of California, Davis, 191 Briggs Hall, One Shields Avenue, Davis, CA, 95616, USA.
authors.forename: Jennifer E
authors.initials: JE
authors.lastname: Gilda
authors.forename: Aldrin V
authors.initials: AV
authors.lastname: Gomes
mesh_terms: D015153:Blotting, Western / Q000379:methods; D011506:Proteins / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D011506:Proteins