Activated peroxidase is an amine-reactive form of horseradish peroxidase (HRP) that provides coupling efficiencies of greater than 95% with antibodies and other proteins.
- 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 1 mg-quantity of activated enzyme is sufficient for reaction with 1 mg of IgG to produce about 0.5 ml 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
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.
Delivery information: Kit contains EZ-Link™ plus activated peroxidase, 5×1 mg, sodium cyanoborohydride solution (5 M), 1×0.5 ml, quenching buffer, 1×25 ml (contains 3 M ethanolamine, pH 9), BupH™ phosphate buffered saline pack, 1 each (results in 0.1 M sodium phosphate, 0.15 M sodium chloride; pH 7.2 when reconstituted with 500 ml of ultrapure water), and BupH carbonate-bicarbonate buffer pack, 1 each (results in 0.2 M carbonate-bicarbonate, pH 9.4 when reconstituted with 500 ml of ultrapure water). This kit is sufficient for five enzyme-labeling reactions, each with up to 1 mg of antibody (IgG).
Packaging: Product is shipped with an ice pack.
Caution: For research use only. Not for use in diagnostic procedures.
