Fluorescein-Conjugated Goat F(ab')2 Fragment to Human Complement C3 is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat F(ab')2 fragment to human complement C3 and buffer salts.
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 susceptible to bacteria infection.
Applications
Fluorescein-Conjugated Goat F(ab')2 Fragment to Human Complement C3 has been successfully used in immunohistochemistry (frozen) applications. It is suitable for use 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. It has also been used in flow cytometry studies. It has been used to assess the binding of complement factors.
Typical Working Concentration
Immunohistochemistry (Frozen) (IHC (F)) 1:50 (dilution). Dilution Recommended: 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immuno Blotting; 1:1,000 - 1:4,000 for ELISA/FIA
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.
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.