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 non-immunoglobulin serum proteins is not present.
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
IgG antibody plays a crucial role in humoral immune responses such as phagocytosis, complement activation, placental transport and cell surface-receptor binding. FITC (fluorescein isothiocyanate), a fluorochrome dye conjugation is useful in localization of antigen-antibody interaction of biological structures like cells or tissues. Anti-cat IgG (whole molecule) − FITC antibody can be used in immunofluorescence study of exfoliative dermatitis in cats. Goat anti-cat IgG antibody is immonospecific for cat IgG. IgG antibody subtype is the most abundant serum immunoglobulins of the immune system. It is secreted by B cells and is found in blood and extracellular fluids and provides protection from infections caused by bacteria, fungi and viruses. Maternal IgG is transferred to fetus through the placenta that is vital for immune defence of the neonate against infections. Anti-Cat IgG (whole molecule)−FITC antibody specifically recognizes cat IgG. Goat anti-cat IgG is conjugated to Fluorescein Isothiocyanate (FITC) in an alkaline reaction, then further purified to remove unbound FITC.
Applications
Fluorescein-Conjugated Goat IgG Fraction To Cat IgG (Whole Molecule) is suitable for use as a reagent in immunofluorescence assays (IFA), cell staining (fluorescent microscopy and cell sorting), tissue staining, and blot immunostaining.
Product Description
Product is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to cat IgG (whole molecule) and buffer salts.
Recommended Use
Reconstitution: 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
Anti-Cat IgG (whole molecule)-FITC antibody may be used for immunofluorescence at a working antibody dilution of 1:32. The antibody was used in indirect fluorescent antibody test (IFAT) to detect Toxoplasma specific antibodies in cat at a dilution of 1:100. Anti-cat IgG (whole molecle)−FITC antibody can be used in direct (1:32) and indirect immunofluorescence (dilution 1:64)