Analysis of Workshop “L” Series Antibodies: Radioimmunobinding and Biochemical Studies

Twenty-one of the Workshop “L” series antibodies (L5 was missing) were received in April, 1984. They were thawed and stored at 8°C throughout the study (April-August, 1984). Aliquots were diluted 1:20 for use in both serological and biochemical assays, the diluent being RPMI 1640 medium containing 5% fetal calf serum (FCS) and 5% horse serum (HS) plus 0.02% sodium azide. All the antibodies were tested for the ability to bind to a selected panel of cells comprising established human leukocyte lines and fresh tissue from cases of leukemia or lymphoma. Those which gave positive results were then examined by “Western Blotting” on solubilized membrane glycoproteins, derived from human leukemic cells, which had been run in polyacrylamide gel electrophoresis (PAGE) and electrotransferred to cellulose nitrate sheets.