How Does the Concentration of the Separating Gel in Western Blot Affect Protein Separation Based on Molecular Weight (kDa)
In Western blot, the concentration of the separating gel determines the resolution of proteins across different molecular weight ranges. Higher gel concentrations provide better resolution for low molecular weight proteins, whereas lower gel concentrations are preferable for high molecular weight proteins. Specifically:
1. Low-concentration gels (e.g., 5%) are commonly used for resolving high molecular weight proteins (>200 kDa).
2. Medium-concentration gels (e.g., 10%-12%) are typically used for proteins ranging from 25 to 200 kDa.
3. High-concentration gels (e.g., 15% or higher) are optimal for resolving low molecular weight proteins (<25 kDa).
Selecting an appropriate gel concentration is essential for achieving efficient protein separation based on molecular weight.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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