Product is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to mouse complement C3 and buffer salts.
Applications
Fluorescein-Conjugated Goat IgG to Mouse Complement C3 is intended for laboratory use to qualitatively stain sections of frozen tissue, 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.
Recommended Use
Reconstitute product with 2.0 mL of deionized or distilled water. Gentle swirling may be used to speed rehydration. Avoid vigorous shaking of the reconstituted material.
Typical Working Concentration
Suggested working dilutions: 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immunoblotting; 1:1,000 - 1:4,000 for ELISA
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 IgG fraction is tested for purity and specificity at 40 mg/mL using immunoelectrophoresis; No trace of albumin is detected. Antibody activity to other serum proteins is not present.