In this paper, AdaBoost algorithm, a popular and effective prediction method, is applied to predict the subcellular locations of Prokaryotic and Eukaryotic Proteins-a dataset derived from SWISSPROT 33.0. Its prediction ability was evaluated by re-substitution test, Leave-One-Out Cross validation (LOOCV) and jackknife test. By comparing its results with some most popular predictors such as Discriminant Function, neural networks, and SVM, we demonstrated that the AdaBoost predictor outperformed these predictors. As a result, we arrive at the conclusion that AdaBoost algorithm could be employed as a robust method to predict subcellular location. An online web server for predicting subcellular location of prokaryotic and eukaryotic proteins is available at http://chemdata.shu.edu.cn/subcell/ .