Rabbit anti-Mouse (Murine) IgM, Fc Specific Antibody (Biotin)

Antigen: IgM, Fc Specific

Clonality: Polyclonal

Host: Rabbit

Reactivity: Mouse (Murine)

Conjugate: Biotin

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

Catalog no.: ABIN458473

Additional Information

Characteristics: Biotin-conjugated IgG fraction of polyclonal rabbit antiserum to mouse IgM, Fc specific

Immunogen: Purified homogenous IgM isolated from mouse serum. Immunization with intact (19S) and split IgM (7S). Freund’s complete adjuvant is used in the first step of the immunization procedure.

Description: The reactivity of the antiserum is directed to the Fc subunit of the IgM molecule which expresses strict isotypic (class) specificity. It does not react with IgG, IgA and IgG/Fab or any non-Ig protein in mouse serum, as tested by immunoelectrophoresis and double radial immunodiffusion. Cross-reactivity Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since Ig of different species frequently share antigenic determinants.

Specificity: Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since Ig of different species frequently share antigenic determinants. of this immunoconjugate has not been tested in detail.

Application Details

Application Notes: In immunocytochemical and immunohistochemical staining of IgM at the cellular and subcellular level of appropriately treated cell and tissue substrates, to demonstrate circulating IgM antibodies in serodiagnostic microbiology and autoimmune diseases, to identify a specific antigen using a reference antibody of mouse origin known to be of the IgM isotype in the middle layer of the indirect test procedure, in non-isotopic assay methodology (e.g. ELISA) to measure IgM 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 1:250, in ELISA and comparable non-precipitating antibody-binding assays between 1:1,000 and 1:5,000. Adsorption: Immunoaffinity adsorbed using insolubilized antigens as required to eliminate antibodies cross-reacting with other components of the immunoglobulin system or reacting with other serum proteins. Special attention is given to the removal of antibodies to common Ig/Fab. 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 4.7. 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 rabbit 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 storage 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