Real lab failures, root causes, and fixes — curated and bilingually annotated by our team.
Western blot shows high background signal with multiple nonspecific bands, obscuring the target protein band and making interpretation difficult.
Western blot shows no visible band for the target protein, or the band intensity is much weaker than expected, indicating little or no protein was eluted from the immunoprecipitation.
Western blot shows strong bands at approximately 50 kDa (heavy chain) and 25 kDa (light chain) that may mask or interfere with the target protein band of interest.
No or very weak target protein signal despite using adequate antibody concentration, suggesting inefficient antibody capture by the beads due to isotype incompatibility.
In co-immunoprecipitation experiments, expected interacting proteins are not detected on Western blot, suggesting protein-protein complexes were disrupted during sample handling.
Inconsistent or absent target protein signal across replicate samples, with visible reduction in bead pellet volume, indicating beads were aspirated during wash or supernatant removal steps.
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