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 F(ab')2 fragments are tested for purity and specificity at a minimum of 20 mg/mL using immunoelectrophoresis. The product is goat F(ab')2 ; no trace of albumin, whole molecule IgG or Fc is detected.
Preparation Method
Antibody and FITC are conjugated under defined conditions to obtain the specified molar ratio (3-6mol/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.
Applications
Fluorescein-conjugated Goat F(AB')2 fragment to Human IgM, μ chain is used as a reagent in immunofluorescence assays (IFA), cell staining (fluorescent microscopy and cell sorting), tissue staining, and blot immunostaining. F(ab')2 fragments are recommended for staining of cells or tissues which contain Fc receptors.
Product Description
Fluorescein-conjugated Goat F(AB')2 fragment to Human IgM, μ chain is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat F(ab')2 fragment to human IgM (5Fc μ) and buffer salts.
Typical Working Concentration
Dilution: 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immuno Blotting; 1:1,000 - 1:4,000 for ELISA/FIA