Why Is the Recommended Protein Load for Western Blot 20–30 μg? How Does Loading Affect Results? Does It Depend on the Purpose?
In Western Blot (WB) experiments, the recommended protein loading amount typically ranges from 20 μg to 30 μg. This range is generally sufficient to produce well-defined bands while preventing overloading, which may cause signal saturation or band smearing. Excessive protein loading can lead to overly intense signals, blurred bands, or even band aggregation. Conversely, insufficient protein loading may result in weak signals, making it challenging to detect the target protein.
Moreover, the optimal protein loading amount depends on the specific objective of the WB analysis. For instance, when detecting low-abundance proteins or conducting quantitative analysis, it may be necessary to adjust the loading amount to achieve the best results.
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