Peng S, Garzon DJ, Marchese M, Klein W, Ginsberg SD, Francis BM, Mount HT, Mufson EJ, Salehi A, Fahnestock M. Decreased brain-derived neurotrophic factor depends on amyloid aggregation state in transgenic mouse models of Alzheimer's disease. J Neurosci. 2009 Jul 22;29(29):9321-9. PubMed Abstract

REAGENTS/MATERIAL:
Western blotting We compared the reaction profiles of three monoclonal antibodies, mouse anti-Aβ (NU-2) (ADDL specific), mouse monoclonal anti-Aβ (6E10) (Signet Laboratories); and mouse monoclonal anti-APPa (22C11) (Millipore), with mouse cortical homogenates. NU-2 strongly recognized 12-mers (Aβ*) in all four strains of AD and Down syndrome mice, demonstrating that these small Aβ oligomers are not responsible for the difference in BDNF levels between these mouse strains. 22C11 strongly recognized a 100 kDa band (soluble APP) in all four strains of AD and Down syndrome mice, demonstrating that the high-molecular-weight band specific to APPNLh and TgCRND8 transgenic mice is not APP. However, NU-2 and 6E10 detected strong signals for the ~115 kDa oligomer in APPNLh and TgCRND8 transgenic mice but very weak signals in both APPswe/PS-1 and Ts65Dn mice, implicating this high-molecular-weight species in BDNF downregulation.
ELISA for Aβ42 and Aβ40 Homogenates were appropriately diluted to equal protein concentrations with sample washing buffer (supplied from the kit) and then assayed for total Aβ40 and Aβ42 using a commercially available sandwich-type ELISA (CRP)