Efficient labelling of antibodies with horseradish peroxidase using cyanuric chloride.

An efficient and mild method for labelling of immunoglobulin G (IgG) with horseradish peroxidase (HRP) using cyanuric chloride (2,4,6-trichloro-1,3,5-triazine, CC) as a bridging molecule is described. The enzyme was treated first with cyanuric chloride to introduce dichloro triazine and after removal of excess reagent, the activated enzyme was mixed with the IgG preparation and incubated to effect linkages with amine groups in the antibody protein. Various amounts of coupling reagent were tested to optimise the conjugation method using commercially available enzyme and affinity-purified sheep IgG antibody preparations to three different test haptens.