The reactivity of the antiserum is directed to the Fc subunit of the IgA molecule which expresses strict isotypic (class) specificity. It does not react with any non-Ig protein in chicken serum, as tested by immunoelectrophoresis and double radial immunodiffusion. In enzyme-immunocytochemical and immunohistochemical staining for the detection of IgA at the cellular and subcellular level by staining of appropriately treated cell and tissue substrates; to demonstrate circulating IgA antibodies in serodiagnostic microbiology and autoimmune diseases; to identify a specific antigen using a reference antibody of chicken origin known to be of the IgA isotype in the middle layer of the indirect test procedure; in non-isotopic assay methodology (e.g. ELISA) to measure IgA in chicken serum or other body fluids. Antisera to IgA do not discriminate between serum IgA (monomeric and dimeric) and higher molecular forms such as secretory IgA.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 1:50 and 1:250; in ELISA and comparable non-precipitating antibody-binding assays between 1:500 and 1:5,000.
Horseradish peroxidase-conjugated IgG fraction of polyclonal Goat antiSerum to Chicken IgA, Fc specific
ELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting
Fields of Interest
Purified normal IgA isolated from pooled chicken serum. FreundÕs complete adjuvant is used in the first step of the immunization procedure.
Horseradish 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.
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 years. Reconstituted at +4¡ C--7 days