Quantitative Western Blot Analysis With Replicate Samples

Quantitative Western blotting (QWB) is a powerful method for assessing changes in protein abundance, but its reliability hinges on thoughtful replication and normalization. This manual offers a comprehensive guide to designing experiments with both technical and biological replicates, ensuring that observed effects are statistically meaningful and biologically relevant. It outlines best practices for normalization using total protein stains, housekeeping proteins, and modification-specific antibodies, and provides step-by-step instructions for calculating fold changes, percent changes, and coefficients of variation. With clear guidance on data interpretation and visualization, including scatterplots and error bars, researchers can distinguish real biological effects from assay noise.

Explore how you can improve the precision and reproducibility of your Western blot data.