Fig. 5.

Ammonium sulphate precipitation and immunoprecipitation of rat brain DDHD2. (A) Ammonium sulphate fractionation of rat brain DDHD2. The soluble fraction of rat brain homogenates (105,000 × g supernatant) was further fractionated by ammonium sulphate precipitation. The same proportion of each sample was loaded onto an SDS-PAGE and analysed by western blotting using an anti-rat DDHD2 antibody. (B) Immunoprecipitation of DDHD2 in rat brain. Forty percent ammonium sulphate precipitate was used for the experiment. Ten micrograms of the anti-DDHD2 or control IgG was used. Aliquots of the samples were loaded onto an SDS-PAGE gel and used for the DG lipase assay. DG lipase activity in the immunoprecipitated beads or in the supernatant was measured. [¹⁴C]SAG was used as a substrate. Data are the means of triplicate measurements. Error bars represent standard error of the mean. Data are representative of three independent experiments. *P < 0.01 for the anti-DDHD2 IgG samples versus control IgG samples.