Western Blotting (WB), Immunoprecipitation (IP), Enzyme Immunoassay (EIA), Dot Blot (DB), Immunodiffusion (ID), Immunofluorescence (IF), Radioimmunoassay (RIA)
Specificity
The reactivity of the antiserum is restricted to lactoperoxidase as tested in immunoelectrophoresis and radial immunodiffusion. Using various antiserum concentrations against fresh bovine milk a single precipitin line is obtained. No precipitation reaction is obtained with normal bovine serum or plasma. Cross-reactivity: The antiserum does not cross-react with any other component of bovine serum. Interspecies cross-reactivity is a normal feature of antibodies, since homologous proteins of different species frequently share antigenic determinants. Cross-reactivity of this antiserum has not been tested in detail, but reactivity with lactoperoxidase from goat and sheep may be expected.
Purification
DEAE-column Chromatography
Immunogen
Lactoperoxidase (EC 1.11.1.7) is one of the major enzymes in bovine milk. The enzyme has a molecular weight of 80,000. It is isolated from pooled bovine milk. Freund’s complete adjuvant is used in the first step of the immunization procedure.
Lactoperoxidase is a natural antibacterial agent in bovine milk, a medium otherwise rich in its ability to support bacterial growth. Milk lactoperoxidase levels vary among species, ranging from little or none in humans to very high concentrations in guinea pigs. Interestingly, lactoperoxidase from milk is identical to the lacrimal and salivary forms of the enzyme. In addition, the enzyme catalyzes the hydrogen peroxide oxidation of iodine and can be employed in labelling proteins with radioiodine.Synonyms: EC=1.11.1.7, LPO, Lactoperoxidase, SAPX, SPO, Salivary peroxidase