SDS–PAGE analysis of the purified rLC.
Coomassie Brilliant Blue staining (A) and western blotting (B) analysis of purified rLC compared to AL-2 Bence Jones protein (AL-2 BJ), purified from urine. Protein samples (5 µg loaded) were resolved on 12% SDS-PAGE run under non reducing and reducing conditions. Western blot analysis was performed using a rabbit anti-human λ LC antiserum as primary antibody (Dako Cytomation, Glostrup, Denmark), revealed by a goat anti-rabbit conjugated with horseradish peroxidase (Dako Cytomation), probed with diaminobenzidine (DAB) substrate. Molecular mass markers are shown in the left lane. The proteins presented the typical dimeric-monomeric species of free LC. Different degrees of protein denaturation are responsible of the minor differences in electrophoretic migration of monoclonal free LC (A), a common finding in SDS PAGE running under denaturing conditions of these proteins.