EZ-Link™ Activated Peroxidase Antibody Labeling Kit

The Thermo Scientific Pierce Activated Peroxidase and Antibody Labeling Kit is designed to prepare and purify HRP-conjugated antibodies with any previously unmodified primary or secondary antibody (IgG).

Features of the Activated Peroxidase and Antibody Labeling Kit:

• 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 1 mg-quantity of activated enzyme in the kit is sufficient for reaction with 0.3 mg of IgG to produce about 0.5 mL of conjugate

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.

For additional information about glutaraldehyde conjugation, see our review of reductive amination and the discussion of applications that follows.