We report a simple and sensitive method for label-free detection of single-stranded DNA-binding protein (SSBP) based on an array of microfabricated cantilevers. The single-stranded DNA (ssDNA) was immobilized on the surface of the sensing cantilevers to detect SSBP, while the reference cantilevers were modified with 6-mercapto-1-hexanol to detect any unwanted cantilever deflection. The differential deflection signals that reveal specific SSBP–ssDNA binding have been found to depend on the SSBP concentration. Using the cantilever array sensor we can detect SSBP in the concentration range from 0.01 to 7μgmL−1. Other proteins, such as thrombin or bovine serum albumin induced no significant deflection of the cantilevers. Our results show the potential for the application of cantilever array sensor system as a powerful tool to detect proteins with high sensitivity and specificity.