AbstractNeurotrauma in the form of traumatic brain injury (TBI) afflicts more Americans annually than Alzheimer's and Parkinson's disease combined, yet few researchers have used neuroproteomics to investigate the underlying complex molecular events that exacerbate TBI. Discussed in this review is the methodology needed to explore the neurotrauma proteome—from the types of samples used to the mass spectrometry identification and quantification techniques available. This neuroproteomics survey presents a framework for large‐scale protein research in neurotrauma, as applied for immediate TBI biomarker discovery and the far‐reaching systems biology understanding of how the brain responds to trauma. Ultimately, knowledge attained through neuroproteomics could lead to clinical diagnostics and therapeutics to lessen the burden of neurotrauma on society. © 2006 Wiley Periodicals, Inc., Mass Spec Rev 25:380–408, 2006