Materials

1. Fluorescein isothiocyanate or tetramethylrhodamine isothiocyanate (see Note 1) dissolved in dimethyl sulfoxide at a concentration of 1 mg/mL.

2. Antibody: this should be the pure IgG fraction or, better, affinity-purified antibody from an antiserum or pure monoclonal antibody (see Chapter 7).

3. 0.1 M Sodium carbonate buffer, pH 9.0.

5. Phosphate-buffered saline (PBS): 20 mM sodium phosphate buffer, pH 7.2, containing 0.15 M NaCl.

6. Sephadex G-25 (Amersham Biosciences, Chalfont St. Giles, UK; see Note 2).

7. Bovine serum albumin (BSA).

8. Sodium azide (Note: sodium azide is toxic).

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