Analysis Note
The total protein is measured using the Biuret procedure with bovine albumin as the standard. To ensure that the product titer falls within the required range, antibody titer is standardized by microtiter plate ELISA with human IgG. The product is tested for purity and specificity at final concentration by immunoelectrophoresis.
Preparation Method
Antibody and highly purified HRP (Rz>3.0) are conjugated under defined conditions to obtain optimally labeled product. Conjugated protein is purified by salt fractionation. The product is dialyzed into 0.02M sodium phosphate, 0.14M sodium chloride, pH 7.3, with 0.01% thimerosal, adjusted to standard titer, filtered through a 0.22 μm filter, vialed and lyophilized.
Background Information
Anti-Human secondary antibodies are affinity-purified antibodies with well-characterized specificity for human immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.
Applications
Peroxidase-conjugated goat IgG fraction to human IgG Fc is used in immunostaining of acetone-fixed frozen and formalin-fixed, paraffin-embedded tissue sections. It is also suitable for use as a reagent in enzyme immunoassays (EIA), cell and tissue staining (for light microscopy), cell and tissue labeling (for electron microscopy), and blot immunostaining. (Note: 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 inherent antibodies of host animals.
Product Description
Peroxidase-conjugated goat IgG fraction to human IgG Fc is the lyophilized powder of horseradish peroxidase (HRP)-conjugated goat IgG fraction to human IgG Fc and buffer salts.
Typical Working Concentration
ELISA (ELISA) 1:10,000; Immunohistochemistry (Frozen) (IHC (F)) 1:500; Western Blot (WB) 1:1000, 1:500 - 1:4,000 for Tissue Staining; 1:1,000 - 1:8,000 for Immuno Blotting; 1:2,000 - 1:16,000 for ELISA/FIA