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
Human IgGs are glycoprotein antibodies that contain two equivalent light chains and a pair of identical heavy chains. IgGs have four distinct isoforms, ranging from IgG1 to IgG4. These antibodies regulate immunological responses to allergy and pathogenic infections. IgGs have also been implicated in complement fixation and autoimmune disorders.
Applications
Fluorescein-Conjugated Goat IgG Fraction To Human IgG Fc is used 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 IgG Fc is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to human IgG Fc 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