Gervais FG, Singaraja R, Xanthoudakis S, Gutekunst CA, Leavitt BR, Metzler M, Hackam AS, Tam J, Vaillancourt JP, Houtzager V, Rasper DM, Roy S, Hayden MR, Nicholson DW. Recruitment and activation of caspase-8 by the Huntingtin-interacting protein Hip-1 and a novel partner Hippi. Nat Cell Biol. 2002 Feb;4(2):95-105. PubMed Abstract

REAGENTS/MATERIAL:

Hippi polyclonal antibodies were produced in the lab. Mouse brain sections were incubated with anti-Hippi antibodies at a concentration of 1:100 and 50 g ml-1 biotin for 48–72 h. Other brain sections were sequentially placed into primary antisera against Hippi (diluted 1:100) or NeuN (dilution 1:50), or Hip-1 (diluted 1:100) or GFAP(dilution 1:200)for 24h at 4°C. Cultured neuronal NT2 cells and primary rat striatal neurons were permeabilized by incubation in 1% Triton-PBS solution for 5 min at room temperature, followed by two washes in PBS supplemented with 10 mM glycine, and were stained for 1 h at 4 °C with affinity-purified rabbit polyclonal anti-Hippi and/or mouse monoclonal anti-Hip-1 antibodies followed by two washes in PBS–glycine buffer.