Tpl2 was immunoprecipitated from cell lysates using 5 μg of antibody per mg of extract (lane 1) or control antibody (lane 2). The immunoprecipitated material was subjected to SDS-PAGE, transferred to a PDVF membrane and immunoblotted with anti-Tpl2 at 5 μg/ml. Binding of the primary antibody was detected using rabbit peroxidase conjugated anti- sheep IgG antibody (1 in 10, 000 dilution, Pierce) followed by enhanced chemiluminescence (ECL, Amersham).