Source of Problem | Possible Cause | Action/ Solution |
---|---|---|
Antibody related
|
Primary or secondary antibody concentrations were too low |
|
Low antigen-antibody binding affinity
|
|
|
Antigen related | Insufficient sample loaded on the gel |
|
Blot has been stripped and reporbed |
|
|
Technique related | Low transfer efficiency |
|
Film exposure time (ECL detection) was too short |
|
|
Buffer related | Non-fat dry milk may mask some antigens |
|
Western Blot Troubleshoot: No Bands | Western Blot Troubleshoot: High Background Signal on the Blot |