Avoiding Housekeeping Protein Detection Saturation

Reliable western blot data require detection of the target and loading control proteins in the linear dynamic range. Many published western blot images show saturated protein bands, especially for the housekeeping proteins (HKP) including ß-actin, ß-tubulin, and GAPDH, indicating that they were not measured in the linear dynamic range. Quantitative analysis based on this type of data is not reliable. This application note describes common ways of avoiding signal saturation for the detection of HKP loading controls.