Goat anti-Mouse (Murine) IgG, IgA and IgM (H&L) Antibody (Biotin)

Antigen: IgG, IgA and IgM (H&L)

Clonality: Polyclonal

Host: Goat

Reactivity: Mouse (Murine)

Conjugate: Biotin

Application: ELISA, Immunohistochemistry (IHC), Immunocytochemistry (ICC)

Catalog no.: ABIN458366

Additional Information

Characteristics: Biotin-conjugated IgG fraction of polyclonal goat antiserum to mouse IgG, IgA and IgM, heavy and light chains

Alternative name: IgG, IgA and IgM (Heavy and Light Chains)

Immunogen: Highly purified IgG and pools of homogenous IgA and IgM isolated from mouse serum. Freund’s complete adjuvant is used in the first step of the immunization procedure.

Description: The reactivity of the antiserum is directed to the major isotypes of the mouse immunoglobulin system (classes and both light chain types) including antibodies to common determinants, to class and to the surface determinants of the common Fab portion, as tested by immunoelectrophoresis and double radial immunodiffusion against pooled serum and purified immunoglobulins. In immunoelectrophoresis and double radial immunodiffusion using various antiserum concentrations against normal mouse plasma and serum, the characteristic IgG, IgA and IgM precipitin lines are obtained.

Specificity: Inter-species cross-reactivity is a normal feature of antibodies to mammalian immunoglobulins, since homologous proteins of different species frequently share antigenic determinants. The degree of cross-reactivity is also dependent on the concentrations of the reactants and the sensitivity of the assay arrangement. This antiserum fraction has been tested for cross-reactivity by double radial immunodiffusion against several species sera with the following results: bovine - cat - chicken - dog - duck - guinea pig + hamster ± horse - human - monkey - pigeon - rabbit - rat + sheep - swine - turkey -

Application Details

Application Notes: In immunocytochemical and immunohistochemical staining of immunoglobulins at the cellular and subcellular level of appropriately treated cell and tissue substrates, to demonstrate circulating antibodies in serodiagnostic microbiology and autoimmune diseases, to identify a specific antigen using a reference antibody of mouse origin in the middle layer of the indirect test procedure, in non-isotopic assay methodology (e.g. ELISA) to measure immunoglobulins in mouse serum or other body fluids. As a second step an avidin or streptavidin conjugate of the user’s 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 for 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. Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required to eliminate antibodies cross-reacting with other serum proteins. The use of insolubilized adsorption antigens prevents the presence of excess adsorbent protein or immune complexes in the antiserum.

Concentration: IgG protein concentration 10 mg/ml. Biotin/IgG protein molar ratio (B/P) is approximately 6.4. No foreign proteins added.

Purification: Hyperimmune antisera with strong precipitating activity are selected for fractionation by salt-precipitation and purification of the IgG fraction by DEAE-chromatography.

Buffer: Biotin-coupled purified hyperimmune goat IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). Preservative: No preservative added, as it may interfere with the antibody activity.

Storage: Lyophilised at +4°C (10 years), reconstituted at or below -20°C (3-5 years), reconstituted at +4°C at least 7 days. The lyophilized conjugate is shipped at ambient temperature and may be stored at +4°C, prolonged stora ge at or below -20°C. It is reconstituted by adding 1 ml sterile di stilled 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.

Restrictions: For Research Use only