Triphosophoinositide (TPI) mixed with bovine serum albumin and Freund's complete adjuvant was injected into the foot pads of a rabbit to prepare anti-TPI antiserum. The antibody titer in the serum was examined by ELISA. 40 days after the initial immunization, the titer of anti-TPI antiserum was found to be 1: 2560. The antibody binding to TPI was specifically inhibited with aminoglycoside antibiotics but not with penicillin. By indirect immunofluorescence technique with anti-TPI and anti-aminoglycoside antisera, we found the different distribution patterns of TPI and aminoglycoside in the tissues. In the kidney, TPI was distributed in the glomerular and stromal vessels, while aminoglycoside was in the proximal tubular cells; in the inner ear, it was distributed in the stria vascularis and spiral osseous lamina, but not in the outer hair cells or type I hair cells, which were stained with antiaminoglycoside antiserum.