Reagents

1. Horseradish peroxidase
2. Sodium periodate, 0.1M, freshly made
3. Ethylene glycol
4. Sephadex G-25
5. Sodium acetate buffer, 0.001M, pH 4.2
6. IgG
7. Carbonate:Bicarbonate Buffer, 1.0 M, pH 9.5
8. Sodium tetraborate (4 mg/ml) freshly made.
10. Stir for 20 minutes, RT
11. Add 6 drops ethylene glycol
12. Stir for 5 minutes, RT
14. Pool colored fractions and divide into two equal parts.
15. To the first part, add 14 mg IgG
16. To the second part, add 10 mg IgG.
17. To both add 1M carbonate buffer, dropwise, to raise the pH to 9.0-9.5
18. Stir for 2 hours at room temperature
19. To both add 0.2 ml sodium tetraborate
20. Incubate for 2 hours at 2-8^oC
22. Dialyze against PBS.

Peroxidase Conjugation by Periodate Method

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