The latex of Calotropis procera is a rich source of proteins that have anti-inflammatory, anti-nociceptive and selective cytotoxic and anti-tumorigenic properties. In this study, two distinct protocols were developed to culture C. procera in vitro in order to obtain active molecules. Soluble proteins of both callus and root cultures were extracted and tested for various activities found in the latex. Anti-inflammatory and anti-nociceptive proteins were present in callus and root extracts; however, these proteins did not possess cytotoxic or anti-tumorigenic activity. Larvicidal proteins were present, but they were not related to others that have been reported previously in latex. This study confirms that tissue culture of C. procera is able to produce therapeutic-grade proteins that have the potential to relieve inflammation and pain associated with inflammatory disorders.