Western Blotting Using In-Gel Protein Labeling as a Normalization Control: Stain-Free Technology
- 1 January 2015
- book chapter
- Published by Springer Science and Business Media LLC in Methods in molecular biology (Clifton, N.J.)
- Vol. 1295, 381-391
- https://doi.org/10.1007/978-1-4939-2550-6_27
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.This publication has 13 references indexed in Scilit:
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