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Housekeeping proteins (HKPs) are routinely used as loading controls for Western blot normalization. Using a Housekeeping Protein (HKP) as an Internal Loading Control The internal loading control is used as an indicator of sample protein loading, to correct for loading variation and confirm that observed changes represent actual differences between samples.įor more normalization related resources, see " Further Reading". In quantitative Western blotting (QWB), normalization mathematically corrects for unavoidable sample-to-sample and lane-to-lane variation by comparing the target protein to an internal loading control.
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This protocol is intended for use with near-infrared fluorescent Western blots. Saturated bands and sample overloading frequently compromise the accuracy of QWB.