The reactivity of the antiserum is directed to the Fc subunit of the IgA molecule, which expresses strict (class) specificity. In immunoelectrophoresis and radial in immunodiffusion using various antiserum concentrations against chicken serum, a single precipitin line has been obtained which shows a reaction of identity with the precipitin lines obtained with the purified IgA used as immunogens. It does not react with IgG, IgG/Fab fragments and IgM or any non-Ig protein in chicken serum, as tested by immunoelectrophoresis and double radial immunodiffusion. In immunocytochemical and immunohistochemical staining of IgA at the cellular and subcellular level 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.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 are usually between 1:10 and 1:40, depending on the method used.
Fluorescein isothiocyanate-conjugated IgG fraction of polyclonal Goat antiSerum to Chicken IgA Fc specific
ELISA,Immunocytochemistry,Immunohistochemistry (frozen),(In)direct immunofluorescence
Fields of Interest
Purified IgA isolated from chicken serum. FreundÕs complete adjuvant is used in the first step of the immunization procedure.
FITC-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.
This immunoconjugate is not species-specific since inter-species cross-reactivity is a normal feature of antisera to immunoglobulins. 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