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Fig. 5. Verification of antisera specificity. Phosphorylated or unphosphorylated samples (10 ng) of the indicated arylalkylamine N-acetyltransferases (AANATs) were resolved by SDS/PAGE, analyzed by Western blot, and immunodetected with the indicated immunopurified serum (ID no.) as a dilution of 1:140 to 1:250: Anti-ovine (o)AANAT_194–207 pS205 (As 5901) (A), anti-rat (r)AANAT_22–37 pT31 (As 3352) (B), anti-oAANAT_1–25 (As 3343 ) (C), and anti-oAANAT_1–207 (As 2819) (D). Similar results were obtained in three experiments. The minor bands seen in A and B represent oAANAT_26–207 generated during thrombin digestion (4). Arrowhead, mobility of a 25-kDa standard. For additional details see Materials and Methods.