SU-8 negative photoresist has been extensively employed in the fabrication of microfluidics and microelectromechanical systems. This is due to its advantages including ease of fabrication using limited equipment, biocompatibility, excellent chemical resistance, and compatibility with silicon processing. In addition, its low Young’s modulus compared to silicon has made it an excellent choice for microcantilever structure development especially for sensing applications. This paper presents a fabrication protocol for the development of thin SU-8 microcantilevers. Factors such as baking temperatures and release methods that influence the fabrication yield of SU-8 microcantilevers are considered. The influence of the baking temperature on the deformation of the SU-8 structure is investigated using the finite element method and verified experimentally. Three release methods are studied including a dry release method using fluorocarbon film, and two wet release methods using OmniCoat sacrificial layer and polymethyl methacrylate (PMMA) sacrificial layer. The wet release method using PMMA sacrificial layer produced the highest yield. An aptasensor is formed using the SU-8 microcantilever, and its deflection measured for thrombin detection.