Fluorescein-Conjugated Goat IgG Fraction To Human IgM

Analysis Note The total protein is measured using the Biuret procedure with bovine albumin as the standard; F/P ratio is calculated using absorbance at 491.5nm. Antibody titer of the IgG fraction is standardized by equivalence-point precipitation. The IgG fraction is tested for purity and specificity at 40 mg/mL using immunoelectrophoresis. The product is mostly goat IgG; no trace of albumin is detected. Preparation Method Antibody and FITC are conjugated under defined conditions to obtain the specified molar ratio (3-6 mol/mol). Conjugated protein is purified by gel filtration. The product is dialyzed into 0.02M sodium phosphate, 0.14M sodium chloride, pH 7.3, with 0.05% sodium azide, adjusted to standard titer, filtered through a 0.22 µm filter, vialed, and lyophilized. Background Information Immunoglobin M (IgM) antibody is prominent in early immune responses to most antigens and is the predominant response in certain antibody responses such as "natural" blood group antibodies. IgM is the major immunoglobin expressed on surface B lymphocytes. Most of the chronic lymphocytic leukemias, non Hodgkin lymphomas and Burkitt's lymphoma bear surface IgM, whereas plasmocytes from Waldenström's disease bear intracytoplasmic IgM. Applications Fluorescein-Conjugated Goat IgG Fraction To Human IgM is use as a reagent in immunofluorescence assays (IFA), cell staining (fluorescent microscopy and cell sorting), tissue staining, and blot immunostaining. (Note: F(ab')2 fragments are recommended for staining of cells or tissues which contain Fc receptors. Affinity purified antibodies are recommended to avoid non-specific binding from the inherent antibodies of host animals). Product Description Fluorescein-Conjugated Goat IgG Fraction To Human IgM is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to human IgM (5Fc µ) and buffer salts. Typical Working Concentration Dilution Reccomondations: 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immuno Blotting; 1:1,000 - 1:4,000 for ELISA/FIA