a
Left panel Western blot analysis showing a Her2 protein band (185 kD) in a positive control lane containing human recombinant Her2 protein or tumor lysates of SK-BR-3, MDA-MB-231, and Daudi cells using either anti-Her2 antibody or culture supernatants from indicated culture conditions; Right panel Western blot analysis showing an EGFR protein band (170 kD) in a positive control lane containing human recombinant EGFR protein or tumor lysates of SK-BR-3, MCF-7, and Daudi cells using either anti-EGFR antibody or culture supernatants from indicated culture conditions. b Shows detection of anti-Her2 antibodies against human recombinant Her2 protein in culture supernatants stimulated with SK-BR-3 (upper panel) or Daudi cells (lower panel) from the indicated culture conditions by ELISA. Herceptin® was used as a positive control and anti-CD20 antibody as a negative control; c Immunostaining of SK-BR-3, MCF-7 or Daudi cells using anti-Her2 antibody, immune serum from a stage IV metastatic breast cancer patient or culture supernatants from indicated culture conditions. Staining with irrelevant controls was negative when Rituxan® was used to stain SK-BR-3 cells or when Herceptin® was used to stain Daudi cells, and d Shows the anti-SK-BR-3 antibody levels by whole cell ELISA after 3-serial absorptions of culture supernatants to SK-BR-3 cells (n = 4)