Affinity isolated antibody is obtained from anti-sheep IgG antiserum by immunospecific purification which removes essentially all donkey serum proteins, including immunoglobulins, which do not specifically bind to sheep IgG. Anti-Sheep IgG is conjugated to alkaline phosphatase by protein cross-linking with 0.2% glutaraldehyde. Specificity of the antiserum is determined by immunoelectrophoresis (IEP), prior to conjugation. By IEP, the antiserum reacts specifically with normal sheep serum and sheep IgG. Identity and purity of the antibody is established by immunoelectrophoresis, prior to conjugation. Electrophoresis of the product followed by diffusion against anti-horse IgG and anti-donkey whole serum results in single arcs of precipitation.
