Studies have shown that aluminum (Al) is the most abundant neurotoxic element on Earth, and is implicated in the pathogenesis of Alzheimer's disease (AD). However, the mechanisms underlying Al-induced neurotoxicity are still largely elusive. Based on affinity analyses with Al and LC-LTQ-MS, we have found that serum albumin, brain CK-B and 14-3-3ζ protein have a high affinity for Al3+, and albumin has a much stronger affinity for Al than transferrin. The normal activity of CK-B, and physiological combination of 14-3-3ζ with tau can be severely perturbed by Al. We anticipate that our assay will provide a new focus concerning the mechanism underlying Al-induced neurotoxicity, and aid the design of strategies to prevent AD and other human diseases related to Al overload.