Clarithromycin antibiotic was labeled with technetium-99m by adding 99mTc to clarithromycin in the presence of SnCl2·2H2O. The radiochemical yield of 99mTc-clarithromycin was determined by chromatographic methods. Gel chromatographic method was investigated for separation of the reaction mixture using different eluents including NaCl and phosphate, citrate, and carbonate buffer solutions. Also, free 99mTcO4- and 99mTc-clarithromycin were efficiently separated by reversed-phase HPLC on RP18 column. The maximum radiochemical yield reached 98 ± 0.2%. 99mTc-clarithromycin is stable for 6 h. Biological distribution of 99mTc-clarithromycin was studied for mice with the infection in the left thigh induced using Staphylococcus aureus. The target-to-nontarget (T/NT) ratio for 99mTc-clarithromycin was found to be 7.33 ± 0.13 at 2 h after intravenous injection, with the subsequent gradual decline. This value is higher than that of commercially available 99mTc-ciprofloxacin. The abscess to normal muscle ratio shows that 99mTc-clarithromycin can be successfully used for infection imaging, allowing differentiation between bacterial infection and sterile inflammation.