The self-association of neopluramycin and the binding of this drug to calf thymus DNA were investigated by using UV-visible spectroscopy. This classical pluramycin antibiotic self-associates in 1:1 ethanol - water mixture. Starting from a simple dimerization model, the molar absorption coefficient of monomer, the molar absorption coefficient of dimer and the dimerization constant were determined. The binding constant of neopluramycin to DNA was determined using Wolfe and Scatchard methods.