Primary Antibodies for Normalization
When performing two-color detection, loading controls and normalization are essential for reliable, precise comparison of protein levels. LICORbio offers primary antibodies for normalization that can be used for two-color normalization when performing multiplex Western blots or for cell-based assay normalization.
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When performing two-color detection, loading controls and normalization are essential for reliable, precise comparison of protein levels.
LICORbio offers nine primary antibodies for normalization – antibodies to actin, COX IV, GAPDH, histone h3, tubulin, and vinculin – that can be used for two-color normalization when performing multiplex Western blots or for cell-based assay normalization, such as for In-Cell Western™ Assays.
- For Western blots, lysate preparation, sample loading, and membrane transfer introduce unavoidable variation. Housekeeping proteins can be used for normalization of protein levels.
- Normalize and correct for uneven loading, using the intensity of the internal control band
- Visually compare protein levels with confidence, even if you don't quantify bands
- Ensure that visually-observed changes in protein levels represent actual change, not artifacts
- Read about types of internal control proteins to use
When lanes are heavily loaded with lysate (>15 µg/lane), a highly expressed internal control protein may not be an appropriate choice for normalization. For quantitative analysis of a low-abundance target protein, a low-abundance internal control is recommended. COX IV is an appropriate choice.
For In-Cell Western™ Assays, a second protein target (such as actin, tubulin, COXIV, or GAPDH) can be used for normalization. Abundance of the normalization target must be unaffected by the cell treatments used.
