Mouse IgM (Fc specific) Rabbit Polyclonal Antibody
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Specifications
Product Data | |
Applications | ELISA, ID, IF, IHC, IP |
Recommended Dilution | ELISA. Immunocytochemistry. (In)direct immunofluorescence Immunohistochemistry on Frozen Sections. Direct immunofluorescence staining of cytoplasmic Ig of appropriately treated cell and tissue substrates; to demonstrate immunoglobulins or specific antibodies in cells and tissues; to identify circulating antibodies in serodiagnostic microbiology and autoimmune diseases. 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. Recommended Working Dilutions: 1/10- 1/80. |
Reactivities | Mouse |
Host | Rabbit |
Isotype | IgG |
Clonality | Polyclonal |
Immunogen | Purified homogenous IgM isolated from Mouse serum. Immunization with intact and split IgM. Freund’s complete adjuvant is used in the first step of the immunization procedure. |
Specificity | 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 purified IgG, IgG/Fab fragments and IgA 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. Cross-reactivity of this immunoconjugate has not been tested in detail. |
Formulation | PBS, pH 7.2 Label: TRITC State: Lyophilized hyperimmune IgG fraction Stabilizer: None Preservative: None, as it may interfere with the antibody activity. Label: Tetramethylrhodamine Isothiocyanate isomer R. It has an orange-red fluorescence. To avoid nonspecific background staining, specially synthesized and exceptionally pure crystalline isomer R has been used instead of the usual racemic mixture. Although its fluorescence efficiency is less than of FITC, TRITC conjugates have the advantage of significantly less photo bleaching. This facilitates their use in quantitative cell-counting procedures. Absorption emission: 554 nm / 573 nm Molar radio: Fluorochrome/IgG protein (F/P) ~ 1.1 |
Reconstitution Method | Restore with 1 ml sterile distilled water. |
Concentration | 10.0 mg/ml |
Purification | Hyperimmune antisera with strong precipitating activity are selected for fractionation by salt precipitation and purification of the IgG fraction by DEAE Chromatography |
Conjugation | TRITC |
Storage | Store lyophilized at 2-8°C for 6 months or at -20°C long term. After reconstitution store the antibody undiluted at 2-8°C for one month or (in aliquots) at -20°C long term. Avoid repeated freezing and thawing. |
Note | 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. Conjugation procedure: A proprietary technique for the binding to TRITC is used, followed by several purification steps to remove free reactants and protein aggregates. After each step activity and specificity are tested in a variety of techniques. The conjugate is lyophilized to assure stability and long shelf life. |
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