The HLA-Bw73 antigen has been characterized by antisera in the Ninth International Histocompatibility Workshop. The International Workshop antibodies 9w245, 9w246, and 9w247 detected HLA-B7 and one or more atingens of this group (HLA-B40, Bw22, Bw42, or Bw48) in addition to HLA-Bw73. We have serologically characterized three additional antibodies, in two family studies, which contain anti-Bw73 (two of the antisera also contain anti-B7 activity). We have performed absorption studies with the three antisera, which indicate that anti-Bw73 activity is removed by HLA-B7 positive lymphocytes in two of the antisera and that, in one case, anti-B7 activity is removed by HLA-Bw73 positive for HLA-B7 negative lymphocytes. The third antiserum is cytotoxicity negative absorption positive for HLA-B7. Neither HLA-B27 positive nor HLA-B8 positive lymphocytes removed any antibody activity. Using one-dimensional isoelectric focusing, unique bands have been characterized for over 30 Caucasian allotypes, including HLA-B7 and HLA-B27. Lymphocytes from two families carrying the HLA-Bw73 antigen were analyzed by isoelectric focusing. These two families show that HLA-Bw73 has a band migrating in the gel very close to HLA-B35 but distant from the cross-reactive group HLA-B7. These studies indicate that HLA antigens which share common epitopes (including those recently characterized, such as HLA-Bw73 and HLA-B7), can be distinguished serologically and by isoelectric focusing.