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Immunogen
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Human J chain is a polypeptide folded within the structure of the polymeric immunoglobulin, resulting in a very limited exposure of J chain antigenic determinants. The antiserum is raised against the isolated and purified J chain. J chains isolated from polymeric IgA and IgM are identical by criteria of composition, peptide maps and antigenicity, Human J chain is distinct from all other chain components of polymeric IgA and IgM. It has a unique primary structure as shown by sequence analyses. Freund's complete adjuvant is used in the first step of the immunization procedure.
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Isotype
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IgG
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Specificity
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Tested in immunoelectrophoresis and radial immunodiffusion (Ouchterlony), this immunoconjugate shows a single precipitation reaction with totally reduced and alkylated polyclonal and monoclonal polymeric IgA, secretory IgA and IgM. A reaction of complete identity is obtained with precipitated highly purified J chain and with the single of precipitation obtained with normal human serum after two hours incubation with 9 M urea and 0.2 M mercaptoethanol at pH 8.6. In a competitive radioimmunoassay no inhibition is obtained with monomeric IgA, polyclonal IgG, reduced and alkylated alpha, mu, gamma, kappa and lambda light chains (less than 1 percent over background)
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Cross-Reactivity (Details)
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The antiserum does not cross-react with any other component of human plasma. Inter-species cross-reactivity is a normal feature of antibodies to plasma proteins since they frequently share antigenic determinants. J chains of many related specie (e.g. mouse, rabbit, dog cat sheep and swine) are known to posses a striking structural homology. of this antiserum has not been tested in detail.
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Characteristics
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Tetramethylrhodamine isothiocyanate-conjugated IgG fraction of polyclonal rabbit antiserum to human J chain of dimeric IgA
Fluorochrome/IgG protein molar ratio (F/P) approximately 2.0. No foreign proteins added.
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Purification
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Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required, to eliminate antibodies reacting with other with other plasma proteins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum. Hyperimmune antisera with strong precipitating activity are selected for fractionation by saltprecipitation and purification of the IgG fraction by DEAE-chromatography.
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