Real lab failures, root causes, and fixes — curated and bilingually annotated by our team.
No bands appear when testing recombinant protein or overexpression lysate, despite expected high protein levels, indicating structural or epitope accessibility issues.
Persistent high background signal related to membrane choice, particularly when using PVDF membranes which generally produce more background than nitrocellulose.
Protein degradation, dephosphorylation, or denaturation occurs when samples are kept above 4°C throughout the protocol, resulting in high background or loss of target bands.
Only sample lanes show faint or absent bands while molecular weight ladder is clearly visible, suggesting issues specific to target protein detection rather than general transfer problems.
Excessive background signal across the membrane obscures specific bands, caused by non-specific antibody binding, inadequate blocking, or improper handling.
All bands on the blot, including the molecular weight ladder, are difficult to see or completely absent, indicating a systemic technical problem rather than target-specific detection failure.
Insufficient lysis strength results in incomplete protein extraction, leading to weak target signal and increased nonspecific bands from partially solubilized proteins.
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