This study developed a practical two-microphone impedance tube method to measure the sound transmission loss of sound isolation materials without the use of an expensive reverberation room or an acoustic intensity probe. In order to evaluate the validation and applicability of the two-microphone impedance tube method, sound transmission losses for several sound isolation materials with different surface density and bending stiffness were measured, and the measured values were compared with the results from the reverberation room method and the theory. From the experimental results, it was found that the accuracy of sound transmission loss obtained by the impedance tube method depends upon the diameter size of the impedance tube (i.e., tested sample size). For sound isolation materials having relatively large bending stiffness such as acryl, wood, and aluminum plates, it was found that the impedance tube method proposed by this study was not valid to measure the sound transmission loss. On the other hand, for sound isolation materials having relatively small bending stiffness such as rubber, polyvinyl, and asphalt sheets, the comparisons of transmission loss between the results from the impedance tube method and the theory showed a good agreement within the range of the frequencies satisfying the normal incidence mass law. Therefore, the two-microphone impedance tube method proposed by this study can be an effective measurement method to evaluate the sound transmission loss for soft sound isolation sheets having relatively small bending stiffness.