Overview:

Affinity purified rabbit immunoglobulins were used to raise goat antibodies specific to the rabbit IgG.The goat immune serum is subsequently affinity purified using immunoglobulins immobilized to agarose.The antibodies are then conjugated to horse radish peroxidase (HRP) by reductive amination in a 2:1 molar ratio. The HRP conjugated secondary antibody can recognize the primary immunoglobulin and give a signal in the presence of a chemiluminescent substrate.The signal can be detected using an electronic imager or autoradiography film (1).

References:

1.Nakane PK, Pierce GB: Enzyme-labeled antibodies: preparation and application for localization of antigens, J Histochem Cytochem 1966, 14:929