Fluorescein-Conjugated Rabbit IgG Fraction to Dog IgG (Whole

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. Rabbit polyclonal anti-Dog IgG (whole molecule)–FITC antiserum is determined to be immunospecific for dog IgG by immunoelectrophoresis versus normal dog serum and dog IgG, prior to conjugation. Second antibodies (secondary antibodies) are antibodies raised against other antibodies, typically from another species, called primary antibodies. Primary antibodies are generated with and bind to specific antigens to create antigen antibody complexes which may be detected in a variety of immunochemical and immunohistochemical configurations involving soluble or insoluble signaling molecules. The immunodetection system depends upon the binding of a signal competent secondary antibody to a primary antibody:antigen complex. The suitability of any secondary antibody to a specific immunodetection application is dependent upon its signal development system configuration. Signal development systems are based upon the chemical conjugation of a signal mediator to the secondary antibody. Major secondary antibody signal systems are color (chromogenic), chemiluminescence or fluorescence based and involve the use of enzymes or high affinity binary systems such as biotin:avidin. Fluorescein isothiocyanate (FITC) is a fluorescein derivative (fluorochrome) used to tag antibodies, including secondary antibodies, for use in fluorescence-based assays and procedures. FITC excites at 495 nm and emits at 521 nm. Fluorescein-Conjugated Rabbit IgG Fraction to Dog IgG (Whole Molecule) used in immunostaining of acetone-fixed frozen and formalin-fixed, paraffin-embedded tissue sections. Suitable for use as a reagent in immunofluorescence assays (IFA), cell staining (fluorescent microscopy and cell sorting), tissue staining, and blot immunostaining. Product is the lyophilized powder of fluorescein-5-isothiocyanate (FITC "Isomer I")-conjugated rabbit IgG fraction to dog 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.