Zerbinatti CV, Wozniak DF, Cirrito J, Cam JA, Osaka H, Bales KR, Zhuo M, Paul SM, Holtzman DM, Bu G. Increased soluble amyloid-beta peptide and memory deficits in amyloid model mice overexpressing the low-density lipoprotein receptor-related protein. Proc Natl Acad Sci U S A. 2004 Jan 27;101(4):1075-80. PubMed Abstract

REAGENTS/MATERIAL:

Mice used were PDAPP (Games, 1995) and mLRP2 (Borchelt, 1996). Double TG mice overexpressing APPV717F and mLRP2 transgenes were obtained by breeding homozygous (+/+) PDAPP mice with heterozygous (+/–) mLRP2 mice that were bred back into the C57BL/6 background for at least five generations.

Brain lysates, pellets and cerebrospinal fluid were used for measurement of soluble/insoluble Ab by sandwich ELISA for human Ab40 (2G3 antibody) and human Ab42 (21F12 antibody). For IHC, Brain sections were stained with a polyclonal antibody to HA epitope (Upstate Biotechnology) to detect mLRP2 or with a polyclonal antibody to full-length LRP to detect both endogenous LRP and mLRP2. A rabbit polyclonal pan-Ab antibody (BioSource International), followed by diaminobenzidine detection, was used to visualize Ab load. Western blots were probed by 6E10 anti-Ab monoclonal antibody (Signet).