A fragment encoding 327 amino acids (1585–1912 in AB257853) of the unc-79 protein was subcloned into the pGEX-4T1 vector (GE Healthcare). The resulting GST-fusion protein was purified and used as an antigen to raise chicken polyclonal antibodies (Aves Antibodies, Tigard, OR). A MBP-fusion to the same 327 amino acid stretch was used for subsequent affinity purification. Whole brain lysates from wild-type and Lwt/Lwt mutant P0 animals were electrophoresed on 4–12% reducing Tris-Acetate gels and transferred to a nitrocellulose membrane (Invitrogen, Carlsbad, CA). After incubation with primary antibody, a goat anti-chicken HRP-conjugated secondary antibody (Aves Antibodies) and ECL reagent (GE Healthcare) was used for detection.
