Beglopoulos V, Sun X, Saura CA, Lemere CA, Kim RD, Shen J. Reduced beta-amyloid production and increased inflammatory responses in presenilin conditional knock-out mice. J Biol Chem. 2004 Nov 5;279(45):46907-14. PubMed Abstract

REAGENTS/MATERIAL:

The generation of PS1 conditional knock-out (PS1 cKO), PS2–/–, and PS cDKO mice has been described previously. Neural progenitor cell (NPC)-specific PS cDKO mice were generated by crossing floxed PS1 mice, nestin-Cre transgenic mice, and PS2–/– mice.

Western blots were incubated with the Saeko antiserum followed by 125I-labeled anti-rabbit antibody (ICN Biomedicals); antibodies against glial fibrillary acidic protein (GFAP) (1:100,000; Sigma), cathepsin S (1:1000; kind gift of Guo-Ping Shi, Boston, MA), or complement component C1q (1:1000; Quidel) and developed with enhanced chemiluminescence (PerkinElmer Life Sciences). Blots were re-probed with an antibody against a-tubulin (1:100,000; Sigma) as a loading control.

For immunohistochemistry antibodies against cathepsin S (1:30, Santa Cruz Biotechnology), complement component C1q (1:100, HyCult Biotechnology), CD45 (1: 1000, Serotec), anti-GFAP (1:500; Sigma)were used.