Immunofluorescence co-localization assay of Plasmodium falciparum clones 3D7 and the isogenic CLAG 9 mutant clone 11E. Antibody raised in mice to CLAG 9 peptide C were used at 1/1,000 dilution in conjunction with antibodies raised in rabbits to the rhoptry protein, RhopH3, at a dilution of 1/600. This was followed by anti-mouse antibodies conjugated to cy2 and anti-rabbit antibodies conjugated to Texas red at a dilution of 1/150. The parasite nuclei were visualized with Hoechst dye (DAPI). The slides were examined with a microscope with filters appropriate for the three fluorescent dyes,and the images merged. A Specific green staining of a schizont of clone 3D7 with the CLAG 9 antiserum, and the specific red staining of the schizont with RhopH3 antiserum. The merging of these two giving a yellow staining indicates that CLAG 9 colocalizes with RhopH3. B Rupturing schizonts of 3D7 showing multiple nuclei of individual merozoites and the co-localization of CLAG 9 and RhopH3 to the rhoptries. C CLAG 9 isogenic mutant clone 11E showing that there was no staining of the rhoptries in this parasite with the CLAG 9 antisera, although there still was specific staining with the RhopH3 antisera.

Gardiner DL, Spielmann T, Dixon MW, Hawthorne PL, Ortega MR, Anderson KL, Skinner-Adams TS, Kemp DJ, Trenholme KR. CLAG 9 is located in the rhoptries of Plasmodium falciparum. Parasitol Res. 2004 93:64-7. Copyright Springer 2010