Crameri A, Biondi E, Kuehnle K, Lütjohann D, Thelen KM, Perga S, Dotti CG, Nitsch RM, Ledesma MD, Mohajeri MH. The role of seladin-1/DHCR24 in cholesterol biosynthesis, APP processing and Abeta generation in vivo. EMBO J. 2006 Jan 25;25(2):432-43. PubMed Abstract

REAGENTS/MATERIAL:

Mice models used in this study were: Seladin-1 heterozygous (from Dr. Feinstein, Quark Biotech Inc) which were bred to obtain seladin-1-deficient mice. In addition the heterozygous seladin-1 mice were bred to SweAPP Tg2576 (SwAPP/seladin-1).

Monoclonal anti-flotillin 1 (clone 18; Transduction Laboratories), monoclonal anti-PrPc POM-1 (kindly provided by Dr A Aguzzi, University of Zurich), monoclonal anti-TfR (clone CD-71; Santa Cruz Biotechnology Inc.), polyclonal anti-human plasminogen (Biogenesis), monoclonal anti-N-terminal APP (clone 22C11; Roche), polyclonal chicken anti-BACE1 (raised against Fc-Asp 2-fusion protein, kindly provided by Dr C Dingwall, GlaxoSmithKline), polyclonal anti-C-terminal APP (Sigma) and monoclonal 6E10 (Signet) antibodies were used for Western blot analysis.

ELISA analysis was performed with a modified sandwich method that detects specifically either Ab40 or Ab42 (Takeda Pharmaceutical Co, Ltd, Japan). Ab was captured with a specific anti-Ab antibody (BNT77). A species ending in residue 40 or 42 were measured using horse radish peroxidase (HRP)-coupled monoclonal antibodies specific for Ab40 (HRP-conjugated BA27) and Ab42 (HRP-conjugated BC05) sequence.