The reactivity of the antiserum is directed to the Fc and Fab subunits of the IgG molecule. It includes some reactivity with other immunoglobulins via the common Fab portion. It does not react with any non-Ig protein in bovine serum, as tested by immunoelectrophoresis and double radial immunodiffusion. In enzyme-immunocytochemical and immunohistochemical staining for the detection of IgG, antigen or antibody, of appropriately treated cell and tissue substrates at the cellular and subcellular level. In non-isotopic assay methodology (e.g. ELISA) to identify and measure a specific IgG in bovine serum or other body fluid. In electron microscopy, since the complex between the conjugated antibody and the antigen also has electron-dense properties.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 for histochemical and cytochemical use are usually between 1:100 and 1:500; in ELISA and comparable non-precipitating antibody-binding assays between 1:1,000 and 1:4,000.
Horseradish peroxidaseÐconjugated IgG fraction of polyclonal Goat antiSerum to Bovine IgG, heavy and light chains
ELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting
Fields of Interest
Purified normal IgG isolated from pooled bovine serum. FreundÕs complete adjuvant is used in the first step of the immunization procedure.
Peroxidase-coupled purified hyperimmune goat IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). No preservative added, as it may interfere with the antibody activity.
Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since Ig of different species frequently share antigenic determinants. Cross-reactivity of this antiSerum has not been tested in detail, however tested in double radial immunodiffusion no reaction has been observed with Human and Swine Serum.
The lyophilized conjugate 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 distilled 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 immunoconjugate Lyophilized at +4¡ C--at least 10 years. Reconstituted at or below -20¡ C--3-5 year. Reconstituted at +4¡ C--7 days