The antiserum does not cross-react with any other human plasma proteins as tested in gel-diffusion techniques. Inter-species cross-reactivity is a normal feature of antibodies to plasma or milk proteins, since homologous proteins of different species frequently share antigenic determinants. of this antiserum has not been tested in detail, however in double radial immunodiffusion a reaction with Rhesus monkey milk has been observed.
Characteristics
Fluorescein isothiocyanate-conjugated IgG fraction of polyclonal goat antiserum to human lactoferrin
Purification
Adsorption: Immunoaffinity adsorbed using insolubilized fractions of human serum and lactoferrin-depleted human milk as required, to eliminate antibodies reacting with other human serum or milk 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 saltprecipitation and purification of the IgG fraction by DEAE-chromatography.
Immunogen
Exocrine organs produce various secretions, each with its characteristic function. Proteins found in secretions may be divided into two groups: those specific for the particular secretion, and plasma proteins independent of the type of exocrine cells. Lactoferrin belongs to the first group. It is an iron containing protein with a molecular weight of 75,000 and it is antigenically different from transferrin. Lactoferrin has a slight anti-microbial action. Originally identified in milk, its presence has also been demonstrated in other secretions as saliva, semen and tears. The immunogen has been isolated from human milk. Freund’s complete adjuvant is used in the first step of the immunization procedure.
As reagent for the direct detection of lactoferrin in human cells, tissues and body fluids in immunofluores-cence, as detection reagent in non-isotopic methodology and solid phase immunochemistry (e.g. ELISA). This immunoconjugate is not pre-diluted. The optimum working dilution of each conjugate should be established by titration before being used. Excess labelled antibody must be avoided because it may cause high unspecific background staining and interfere with the specific signal. Working dilutions are usually between 1:20 and 1:80.
Restrictions
For Research Use only
Format
Lyophilized
Concentration
IgG protein concentration 10 mg/ml. Fluorescein/IgG protein molar ratio (F/P) approximately 1.6. No foreign proteins added.
Buffer
Fluorochrome-coupled purified hyperimmune IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2)
Tested in immunoelectrophoresis against human milk a single precipitin line is obtained. The antiserum does not react with any other protein component of human serum or plasma