IgE, Fc Specific antibody

Antigen: IgE, Fc Specific

Clonality: Polyclonal

Host: Pig

Reactivity: Human

Application: ELISA, Immunofluorescence (IF), Immunocytochemistry (ICC)

Catalog no.: ABIN458245

Additional Information

Characteristics: Purified IgG fraction of polyclonal swine antiserum to human IgE, Fc specific

Immunogen: Purified homogenous IgE isolated from human serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.

Description: The reactivity of the antiserum is directed to the Fc subunits of the IgE molecule which represents strict isotype (class) specificity. In immunoelectrophoresis and double radial immunodiffusion, using various antiserum concentrations against appropriate concentrations of IgE, a single precipitin line is obtained which shows a reaction of full identity with the precipitin line obtained with the used immunogen. It does not react with human IgG, including all subclasses, IgA, IgM and IgG/Fab or any non-Ig protein in human serum. Cross-reactivity Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since immuno-globulins of different species frequently share antigenic determinants.

Specificity: Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since immuno-globulins of different species frequently share antigenic determinants. of this antiserum has not been tested in detail because of the low level of IgE in most species. Physicochemical characteristic IgG protein concentration 10 mg/ml. No foreign proteins added. Performance testing Isotype specificity and quantitative specific recognition ability are further evaluated at the high level of sensitivity by direct singe and simultaneous double staining of different types of human cells, using counterstaining with reference conjugates with a different marker. The immunoconjugate is further tested in ELISA-type assays.

Application Details

Application Notes: As unlabelled secondary antibody for indirect detection of IgE in human cell, tissue substrates and body fluids in immunofluorescence and immunoenzyme assay methods, for the production of immunoconjugates with a selected marker, to prepare insoluble immunoaffinity adsorbents by coupling to an artificial carrier, as catching or detecting antibody reagent in non-isotopic assay methodology (e.g. ELISA) to identify and measure IgE in human serum or other body fluid. When applied in any immunocytochemical or histochemical staining procedure or solid phase coupling technique, the optimum concentration of the IgG preparation should be established. Working dilutions for histochemical and cytochemical use are usually between 1:100 and 1:250, in ELISA and comparable non-precipitating antibody-binding assays between 1:500 and 1:5,000. Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required to eliminate antibody activity reacting with other components of the immunoglobulin system and to any other serum protein. Special attention is given to the elimination of antibodies to the common Fab portion of immunoglobulins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum.

Purification: Hyperimmune antisera with strong precipitating activity are selected for fractionation by salt-precipitation and purification of the IgG fraction by DEAE-chromatography.

Buffer: Purified hyperimmune swine IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). Preservative: No preservative added, as it may interfere with the antibody activity.

Storage: Lyophilised at +4°C (10 years), reconstituted at or below -20°C (3-5 years), reconstituted at +4°C at least 7 days. The lyophilized IgG fraction is shipped at ambient temperature and may be stored at +4°C, prolonged storage at or below -20°C. It is reconstituted by adding 1 ml sterile di stilled water, spun down to remove insoluble particles, divided into small aliquots, frozen and stored at or below -20°C. Prior to use, an aliquot is thawed slowly at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C , not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the product.

Restrictions: For Research Use only