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For example, the anti-CD20 rituximab induces cell death by binding to surface receptors, resulting in a signaling cascade that leads to apoptosis ( Clostridium difficile toxin B ( The tremendous progress in mAb discovery began in 1975, when Khler and Milstein reported in vitro screening and production of murine mAbs from hybridomas ( To overcome the immunogenicity and reduced effector function of murine mAbs, chimeric antibodies (mousehuman) were next developed by fusing the antigen-specific variable domain of a murine mAb with the constant domains of a human mAb

The anti-human primary antibodies used were: anti-CD31/PECAM-1 (Rabbit polyclonal 1:100, Novus) and anti-fibronectin (Mouse monoclonal 1:3000, Sigma-Merck) followed by fluorophore-conjugated secondary Abs: goat anti-rabbit Alexa Fluor 555 (Invitrogen) and goat anti-mouse Alexa Fluor 488 (Invitrogen) both diluted 1:2000