The reactivity of the antiserum is directed to the Fc and Fab subunits of the IgG molecule. It includes a certain degree of reactivity with other immunoglobulins via the common Fab portion. The reactivity against Fab of IgG is less pronounced than in the corresponding antiserum produced in rabbit. It does not react with any non-Ig protein in bovine serum, as tested by immunoelectrophoresis and double radial immunodiffusion. In immunocytochemical and immunohistochemical staining to identify and measure IgG, antigen or antibody, at the cellular and subcellular level by staining of appropriately treated cell and tissue substrates, and to demonstrate circulating antibodies in serodiagnostic microbiology and autoimmune diseases; to identify a specific antigen or immune complex using a reference antibody of bovine origin in the middle layer of the indirect test procedure. As a second step an avidin or streptavidin conjugate of the userÕs choice has to be used.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 fro histochemical and cytochemical use are usually between 1:100 and 1:500; in ELISA and comparable non-precipitating antibody-binding assays between 1:2,000 and 1:10,000.
BiotinÐconjugated 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.
Biotin-coupled purified hyperimmune goat IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). No preservative added
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.
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 in the dark 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 years. Reconstituted at +4¡ C--7 days