Methods

1. Add 0.5 mg of IgG antibody in 100 |L of PBS to 1.5 mg of alkaline phosphatase.

2. Add 5% glutaraldehyde (approx. 10 |L) to give a final concentration of 0.2% (v/v), and stir the mixture for 2 h at room temperature.

3. Dilute the mixture to 1 mL with PBS and dialyze against PBS (2 L) at 4°C overnight (see Note 1).

4. Dilute the solution to 10 mL with 50 ml Tris-HCl buffer, pH 7.5, containing 1 mM MgCl2, 0.02% NaN3, and 2% BSA (see Notes 2-4).

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