Aldehyde-activated for gentle, high-efficiency conjugation.
Thermo Scientific Pierce Plus Activated Peroxidase is an amine-reactive form of horseradish peroxidase (HRP) that provides coupling efficiencies of greater than 95% with antibodies and other proteins.
Plus Activated Peroxidase is horseradish peroxidase (HRP) enzyme, whose native carbohydrates (sugars) have been gently oxidized with periodate to produce amine-reactive aldehyde groups. These carbonyls of Plus Activated Peroxidase spontaneously and efficiently crosslink with primary amines on antibody or other proteins. The method is more effective than other amine-reactive chemistries, such as glutaraldehyde coupling, which often causes polymerization and a greater degree of conjugate inactivation. The pre-activated enzyme eliminates the difficulties inherent in preparing and validating activated peroxidase from scratch. The kit provides the enzyme, accessory reagents and protocol to easily produce high-performance conjugates for Western blotting, ELISA and other detection techniques.
Activated HRP – periodate-treated, aldehyde-activated horseradish peroxidase, ready for conjugation to antibodies and other proteins at sites of primary amines (e.g., lysines)
Permanent conjugation – reacts efficiently (95%) with primary amines to form covalent amide bonds upon treatment with sodium cyanoborohydride (included in kit)
High activity HRP – enzyme activity is 120 to 200 units/mg; lyophilized, activated enzyme is stable for at least 12 months at -20°C
Convenient quantities – each 1mg-quantity of activated enzyme is sufficient for reaction with 1mg of IgG to produce about 0.5mL of conjugate
Customizable – vary the molar ratios, reaction buffer and pH, and other parameters to acheive conjugates with different levels of HRP incorporation and activity
Conjugation scheme for the Plus Activated Peroxidase. In this diagram, the activated HRP is represented by the blue (P) and the antibody by the yellow (P). Antibodies (IgG) are about three times the size of HRP and contain numerous primary amines. The default kit procedure typically results in 1 to 3 HRP molecules per IgG molecule. For additional details about this reaction chemistry, see our review of Reductive Amination.
Imagawa, M., et al. (1982). Characteristics and evaluation of antibody- horseradish peroxidase conjugates prepared by using a maleimide compound, glutaraldehyde and periodate. J. Appl. Biochem.4, 41-57.