Clinical studies often produce fresh tissue samples, which ideally should be immediately snap frozen for storage and subsequent analysis. However, this is often not practically possible, and there is inevitably a time period during which the sample is stored on ice. The delay in freezing may allow endogenous degradation of proteins to occur, affecting 2-D gel protein profiles. This study aims to investigate the type and extent of this degradation by examining how the time-to-freezing delay alters prostatic tissue protein profile. The prostate carcinoma-3 cell line (PC-3), prostate cancer xenografts and canine prostate were used with fluorescence 2-D DIGE to assess protein degradation. It was found that 30-min processing time had minimal effects on the protein profile. Longer delays had little visible effect, but subtle alterations in protein profile began to accumulate as time increased. These data support the practice of completing tissue processing as rapidly as possible, and indicate that short processing times do not notably perturb the 2-D gel spot pattern from prostatic tissue.