Thermoluminescence (TL) measurements in Tl2InGaSe4-layered single crystals have been carried out in the temperature range of 10-200 K at various heating rates (0.2-1.0 K s(-1)) to get information about the characteristics of traps. Two TL overlapping glow peaks related to defect levels have been clearly observed. Thermal cleaning procedure was applied to the glow curves to separate overlapped peaks. Initial rise, peak shape, and heating rate methods were used to calculate the activation energies of the revealed traps. The energy values of 5 and 28 meV were evaluated for the peaks observed at low and high temperatures, respectively. Moreover, heating rate dependence and traps distribution analysis were also investigated on the curve obtained after thermal cleaning. The activation energies of the distributed trapping centers were found to be increasing from 29 to 151 meV with increasing the illumination temperature from 42 to 80 K.