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.
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.
Immunoglobulin G (IgG) is a glycoprotein antibody that regulates immune responses such as phagocytosis and is also involved in the development of autoimmune diseases. Mouse IgGs have four distinct isotypes, namely IgG1, IgG2a, IgG2b, and IgG3. IgG1 regulates complement fixation in mice. Anti-Mouse IgG (whole molecule)-FITC antibody reacts with normal mouse serum and all mouse IgGs.
Fluorescein-Conjugated Goat IgG Fraction to Mouse IgG (Whole Molecule) is suitable for use as a reagent in immunofluorescence assays (IFA), cell staining (fluorescent microscopy and cell sorting), immunostaining of acetone-fixed frozen and formalin-fixed, paraffin-embedded tissue sections, and blot immunostaining. 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 is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat IgG fraction to mouse IgG (whole molecule) and buffer salts.
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-Mouse IgG (whole molecule)-FITC antibody is suitable for use in immunohistochemistry (1:200) and immunofluorescence (1:100).