Figure 1. Schematic of organelle purification and analysis using a monoclonal antibody approach.

A mixed fraction of organelles was purified from N. caninum by disruption of extracellular parasites in the French Press, and isolation of organelles by differential centrifugation and separation on a Percoll gradient. The mixed organellar fraction residing just above the rhoptry/dense granule band was collected and used to immunize a single mouse. Hybridomas were produced and the resulting panel of monoclonal antibodies was screened by IFA. Selected antigens were identified by immunoprecipitation and mass spectrometry.