The tremendous growth in wireless communication needs a compact, wideband, high gain, low-cost antennas for transmitting and receiving signals. This paper focused on the design of rectangular microstrip antenna for ISM Band with a resonant frequency of 2.45GHz. It also examines how the dielectric constant and thickness of substrates affecting the performance of antennas. Four different substrates FR4, Alumina, PTFE, and RT Duroid 5880 with 1.6mm and 0.8 mm thickness are considered in antenna performance comparison. The work is verified up to the simulation level in Advanced Design System (ADS) software. The S parameter simulation results are presented to compare the performance of antenna for various substrate materials.