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 mammalian serum proteins, since homologous proteins of different species frequently share antigenic determinants. of this antiserum has not been tested in detail.
Characteristics
Precipitating polyclonal rabbit antiserum to human alpha-foetoprotein
Purification
Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required, to eliminate antibodies reacting with other human serum proteins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum.
Immunogen
Purified human alpha-foetoprotein is isolated from pooled amniotic fluid and hepatoma serum, using a proprietary multiple step method. Several foetoproteins have been demonstrated in human foetal serum and amniotic fluid. One of the best studied is the alpha-foetoprotein. It cab easily be identified immunochemically because of its distinct antigenicity. In electrophoresis its relative mobility is between the albumin and alpha-1 globulin fractions. Its molecular weight is 64,000, the carbohydrate content is relatively low. The concentration of alpha-foetoprotein gradually decreases during foetal growth. It is not found in normal adult serum, but reappears in most human developing hepatoma. In pooled foetal/newborn serum a concentration of 1 mg/ml has been measured. Freund’s complete adjuvant is used in the first step of the immunization procedure.
In precipitating techniques as immunoelectrophoresis and single and double radial immunodiffusion (Mancini, Ouchterlony) to identify the presence of alpha-foetoprotein or to determine its concentration.
Restrictions
For Research Use only
Format
Lyophilized
Concentration
Total protein and IgG concentrations in the antiserum are comparable to those of pooled normal rabbit serum. No foreign proteins added. Antibody titre: Precipitin titre 1:64 when tested against pooled human amniotic fluid in agar-block immunodiffusion tit
The lyophilized antiserum 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 antiserum. Diluted antiserum should be stored at +4°C, not ref rozen, and preferably used the same day.
The reactivity of the antiserum is restricted to alpha-foetoprotein. In immunoelectrophoresis and radial immunodiffusion, using various antiserum concentrations against amniotic fluid, hepatoma sera and purified alpha-foetoprotein a single precipitin line is obtained. No precipitation reaction is obtained with normal human serum