Why Is the Protein Marker Not Visible on the Membrane After Western Blot Transfer?
This issue pertains to a common occurrence in Western blot experiments. When the protein marker (molecular weight marker) is not visible on the membrane, several factors may be responsible. The following four points are potential primary causes:
1. Selection of Protein Marker
The choice of marker should be based on the molecular weight of the target protein. If the marker's molecular weight is too close to that of the target protein, or if it falls outside the target protein's range, the marker may not be visible during electrophoresis and transfer.
2. Selection of Gel and Blotting Conditions
The parameters used for electrophoresis and transfer (such as voltage, time, temperature, etc.) should correspond to the marker and target protein. Inappropriate parameter settings can lead to the marker not being detected.
3. Chromogenic Reagent
The chromogenic reagent used may affect the marker's visibility. Improper selection, excessive development time, or incomplete coverage of the sample can result in the marker's color fading or being undetectable.
4. Loss or Degradation of the Marker
This may be due to operational issues during sample preparation, such as insufficient resolubilization, inappropriate storage conditions leading to marker degradation, or loss during the transfer process.
These reasons are not exhaustive; other factors may also be involved. A detailed analysis of the experimental steps, optimization of conditions, and consideration of replacing the marker are recommended to address this issue.
MtoZ Biolabs, an integrated chromatography and mass spectrometry (MS) services provider.
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