Sample & Protein
Protein degradation, low loading amount, or extremely low basal expression of target protein.
Gel Preparation & Transfer
Mismatched separation gel concentration; insufficient transfer current or time, resulting in failure of protein transfer onto membrane.
Antibody Incubation
Excessive blocking masking antigen epitopes; invalid primary antibody, low antibody concentration or insufficient incubation time; mismatched host species of secondary antibody.
Chemiluminescence Detection
Expired luminescent substrate solution; too short exposure time to capture signals.