Invitrogen™ Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 647

Donkey Polyclonal Secondary Antibody

To minimize cross-reactivity, the donkey anti-rabbit IgG whole antibodies have been cross-adsorbed against IgG from bovine, goat, chicken, guinea pig, hamster, horse, sheep, mouse, rat, and human. Cross-adsorption or pre-adsorption is a purification step to increase specificity of the antibody resulting in less background staining and cross-reactivity. The secondary antibody solution is passed through a column matrix containing immobilized serum proteins from potentially cross-reactive species. Only the nonspecific-binding secondary antibodies are captured in the column, and the highly specific secondaries flow through. Further passages through additional columns result in highly cross-adsorbed preparations of secondary antibody. The benefits of these extra steps are apparent in multiplexing/multicolor-staining experiments where there is potential cross-reactivity with other primary antibodies or in tissue/cell fluorescent staining experiments where there may be the presence of endogenous immunoglobulins. Using conjugate solutions: Centrifuge the protein conjugate solution briefly in a microcentrifuge before use; add only the supernatant to the experiment. This step will help eliminate any protein aggregates that may have formed during storage, thereby reducing nonspecific background staining. Because staining protocols vary with application, the appropriate dilution of antibody should be determined empirically.