The antiserum does not cross-react with any other component of bovine serum. Inter-species cross-reactivity is a normal feature of antibodies, since homologous proteins of different species frequently share antigenic determinants. of this antiserum has not been tested in detail, but reactivity with lactoperoxidase from goat and sheep may be expected.
Characteristics
Biotin-conjugated IgG fraction of polyclonal rabbit antiserum to bovine lactoperoxidase
Purification
Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required, to eliminate antibodies cross reacting with other components of the immunoglobulin system or reacting with other serum proteins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the anti-serum. Hyperimmune antisera with strong precipitating activity are selected for fractionation by salt-precipitation and purification of the IgG fraction by DEAE-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 sis isolated from pooled bovine milk. Freund’s complete adjuvant is used in the first step of the immunization procedure.
LPO
Reactivity: Human
ELISA
Host: Rabbit
Polyclonal
unconjugated
Application Notes
In immunocytochemical and immunohistochemical staining to identify and measure lactoperoxidase at the cellular and subcellular level by staining of appropriately treated cell and tissue substrates. As a second step an avidin or streptavidin conjugate of the user’s choice has to be used. Working dilutions for histochemical use are usually between 1:100 and 1:250, in ELISA and comparable non-precipitating antibody-binding assays between 1:1,000 and 1:5,000.
Restrictions
For Research Use only
Format
Lyophilized
Concentration
Physicochemical characteristic IgG protein concentration 10 mg/ml. Biotin/IgG protein molar ratio (B/P) approximately 8.9. No foreign proteins added.
Buffer
Biotin-coupled purified hyperimmune rabbit IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2).
Preservative
Without preservative
Storage
4 °C/-20 °C
Storage Comment
The lyophilized immunoconjugate is shipped at ambient temperature and may be stored at +4°C, prolonged storage at or below -20°C. Reconstitute the lyophilized antiserum by adding 1 ml sterile distilled water. Dilutions may be prepared by adding phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the immunoconjugate. Diluted antiserum should be stored at +4°C, not refrozen, and preferably used the same day.
The reactivity of the antiserum is restricted to lactoperoxidase as tested in immuno-electrophoresis 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