Abstract: The present method relates to quantification of prognostic and predictive biomarkers of the PDK/AKT/mTOR pathway, such as GSK3?, S6, CREB, PTEN, AKT and mTOR, using AQUA® analysis to estimate both patient risk and benefit of treatment to patients diagnosed with glioblastoma. Unlike traditional IHC, the AQUA® system is objective and produces quantitative in situ protein expression data on a continuous scale. Taking advantage of the power of the AQUA system, the present method provides a highly robust and standardized diagnostic assays that can be used in the clinical setting to provide physicians with reliable prognostic and predictive information. Glioblastoma multiform (GBM) remains one of the most aggressive human cancers, and biomarkers that provide prognostic and predictive information would be extremely valuable to both the physician and the patient. A patient's risk may be determined using the prognostic biomarkers of the present method.