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
Complement component C3 plays a central role in the activation of complement system. Its activation is required for both classical and alternative complement activation pathways. People with C3 deficiency are susceptable to bacteria infection.
Applications
Fluorescein-conjugated Goat IgG Fraction to Human Complement C3 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). It is also used for Immunohistochemistry techniques at a dilution of 1:50.
Product Description
Fluorescein-conjugated Goat IgG Fraction to Human Complement C3 is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to human complement C3 and buffer salts.
Typical Working Concentration
Dilution Recommendations: 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immuno Blotting; 1:1,000 - 1:4,000 for ELISA/FIA