The Thermo Scientific Pierce Activated Peroxidase and Antibody Labeling Kit are designed to prepare and purify HRP-conjugated antibodies with any previously unmodified primary or secondary antibody (IgG).
Activated Peroxidase is HRP enzyme that has been activated with glutaraldehyde to contain reactive groups that will spontaneously conjugate with amines on an antibody (or other protein). Incubation of this Activated Peroxidase with IgG in sodium carbonate buffer (pH 9.5) results in formation of a conjugate where both the enzyme activity of HRP and the antigen-binding activity of IgG are preserved. The Antibody Labeling Kit includes the Activated Peroxidase and a Protein A/G column and buffers to purify the antibody conjugate from excess, unreacted HRP enzyme.
Activated HRP – glutaraldehyde-activated horseradish peroxidase, ready for conjugation to antibodies and other proteins at sites of primary amines (e.g., lysines)
Permanent conjugation – reacts with primary amines to form covalent amide bonds; no reduction step is necessary to secure the linkage
High activity HRP – enzyme activity is greater than 200 units/mg; lyophilized, activated enzyme is stable for at least 12 months at -20°C
Convenient quantities – each 1mg-quantity of activated enzyme in the kit is sufficient for reaction with 0.3mg of IgG to produce about 0.5mL of conjugate
For additional information about glutaraldehyde conjugation, see our review of Reductive Amination and the discussion of applications that follows.
Imagawa, M., et al. (1982) Characteristics and evaluation of antibody horseradish peroxidase conjugates prepared by using a maleimide compound, gluteraldehyde and periodate. J. Appl. Biochem.4, 41-57.