Isolation and analysis of AC133 protein. (A) Lanes 1 and 2, silver-stained gel of purified AC133 antigen prepared for protein sequence analysis. (B) Lanes 1 and 2, Western blot of partially purified AC133 antigen using AC133 as the primary antibody with an anti-mouse IgG-HRP conjugate as secondary antibody. Secondary antibody was detected by chemiluminescence with the Super-Signal system (Pierce). (C) Endoglyconase treatment of partially purified AC133 antigen. Lane 1, 0 U PNGase F; lane 2, 0.002 U; lane 3, 0.01; lane 4, 0.02 U; lane 5, 0.1 U. AC133 antigen loses approximately 20 to 25 kD upon N-linked deglycosylation.