We have successfully observed two-photon above-threshold ionization in rare gas atoms (Ar, Xe, and He) by the fifth harmonic (25 eV photon energy) of a KrF laser. Use of the energy-resolved photoelectron counting system together with our laser, providing strong 25 eV radiation at 40-100 Hz, enabled us to detect the very weak single-color two-photon above-threshold ionization signals. Experimental data are in good agreement with our theoretical calculations newly developed along the line of multichannel quantum defect theory.