Sheep IgG (Fc specific) Rabbit Polyclonal Antibody
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Specifications
Product Data | |
Applications | ELISA, IF, IHC, WB |
Recommended Dilution | ELISA. Dot blot. Immunoblotting. Immunocytochemistry. Immunohistochemistry on Paraffin Sections. Can be used In immunocytochemical and immunohistochemical staining of IgG at the cellular and subcellular level of appropriately treated cell and tissue substrates; to demonstrate circulating IgG antibodies in serodiagnostic microbiology and autoimmune diseases; to identify a specific antigen using a reference antibody of sheep origin known to be of the IgG isotype in the middle layer of the indirect test procedure; in non-isotopic assay methodology (e.g. ELISA) to measure IgG in sheep 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. Recommended Working Dilutions: Histochemistry and Cytochemical Use: 1/100-1/500. ELISA and comparable non-precipitating antibody-binding assays: 1/1,000-1/10.000. |
Reactivities | Sheep |
Host | Rabbit |
Isotype | IgG |
Clonality | Polyclonal |
Immunogen | Purified normal IgG isolated from pooled sheep serum. 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 IgG molecule which expresses strict isotypic (class) specificity. In immunoelectrophoresis and double radial immunodiffusion using various antiserum concentrations against normal sheep plasma and serum, a single characteristic precipitin line is obtained which shows a reaction of identity with the precipitin line obtained with purified IgG. It does not react with IgM, IgA and IgG/Fab or any non-Ig protein in sheep serum. 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 antiserum has net been tested in detail; however a strong reaction has been observed with goat IgG. |
Formulation | PBS, pH 7.2 Label: Biotin State: Lyophilized purified hyperimmune IgG fraction Stabilizer: None Preservative: None, as it may interfere with the antibody activity. Label: N-Hydroxysuccinimidobiotin Molar radio: Biotin/IgG ~4.9 |
Reconstitution Method | Restore with 1 ml sterile distilled water. |
Concentration | 10.0 mg/ml |
Purification | Hyperimmune antisera with strong activity are selected for fractionation by saltprecipitation and purification of the IgG fraction by DEAE-chromatography. |
Conjugation | Biotin |
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: Conjugation is carried out using a proprietary technique for the binding to biotin, followed by several purification steps. After each step activity and specificity are tested in a variety of techniques. The conjugate is lyophilized to assure stability and long shelf life. No foreign protein has been added. |
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