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. The product is goat F(ab')2; no trace of albumin, whole molecule IgG or Fc 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 and vialed.
Background Information
Human IgGs are glycoprotein antibodies that contain two equivalent light chains and a pair of identical heavy chains. IgGs have four distinct isoforms, ranging from IgG1 to IgG4. These antibodies regulate immunological responses to allergy and pathogenic infections. IgGs have also been implicated in complement fixation and autoimmune disorders.
Applications
Fluorescein-conjugated goat F(ab')2 fragment to human IgG 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. (Note: Affinity purified antibodies and their F(ab')2 fragments are recommended to avoid non-specific binding from the inherent antibodies of host animals. The antibody may also be used in FACS-based assays (1:200).
Product Description
Fluorescein-conjugated Goat F(ab')2 fragment to Human IgG is the fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated goat F(ab')2 fragment to human IgG (whole molecule) and buffer salts.
Typical Working Concentration
Dilution Recommendations: Direct immunofluorescence (1:16); ANA-indirect immunofluorescence (1:32 or 1:64); FACS-based assays (1:200); 1:100 - 1:400 for Tissue Staining; 1:250 - 1:1,000 for Immuno Blotting; 1:1,000 - 1:4,000 for ELISA/FIA