Fig. 4

Identification of valine amide antibodies in mice. Animals defined as R1–R7 (female BALB/c mice, 8–9 weeks, n = 7) were vaccinated with an initial subcutaneous injection with a 1:1 emulsion of 50 µg of keyhole limpet hemocyanin (KLH)-Val-CONH₂ immunoconjugate and complete Freund’s adjuvant, followed by booster injections with incomplete Freund’s adjuvant. Sera were collected and used to identify high-titer valine amide antisera (VAA) in the vaccinated animals using a standard ELISA. The wells were coated with synthetic immunoconjugates of bovine serum albumin (BSA)-[Tyr₀]-Gly_2–4-Val-CONH₂ or BSA-RGDV-COO. The BSA-RGDV-COO⁻-coated wells displayed no significant signal (data not shown, see text for additional details). Assays were performed in triplicate using two serial dilutions of mouse antisera (1:100; 1:1000). The ordinate describes the immunoreactive signals detected as absorbance at 490 nm (A₄₉₀) in the assay. The values are expressed as the mean ± SEM