Hypoxia within solid tumors decreases therapeutic efficacy, and identification of hypoxia markers may influence the choice of therapeutic modality. Here, we used a proteomic approach to identify hypoxia-regulated proteins and validated their use as endogenous indicators of tumor hypoxia. Using two-dimensional gel electrophoresis and PowerBlot (antibody-based array), we identified a group of 20 proteins that are increased >1.5-fold during hypoxia. The majority of these proteins such as IB kinase  (IKK), MKK3b, highly expressed in cancer (HEC), density-regulated protein 1, P150 glued , nuclear transport factor 2, binder of ARL 2, Paxillin, and transcription termination factor I have not been previously reported to be hypoxia inducible. The increase in these proteins under hypoxia was mediated through posttranscriptional mechanisms. We additionally characterized the role of IKK, a regulator of the nuclear factor-B transcription factor, during hypoxia. We demonstrated that IKK mediates cell survival during hypoxia and is induced in a variety of squamous cell carcinoma cell lines. Furthermore, we showed that IKK expression from tumor specimens correlated with tumor oxygenation in patients with head and neck squamous cell carcinomas. These data suggest that IKK is a novel endogenous marker of tumor hypoxia and may represent a new target for anticancer therapy.