Materials

1. Horseradish peroxidase, 1000 U/mg or greater (with 2,2'-azino-bis-[3-ethylbenzthiazoline-6-sulfonic acid] as substrate; see Note 1).

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 periodate.

4. 1 mM Sodium acetate buffer, pH 4.4.

5. 10 mM Sodium carbonate buffer, pH 9.5.

6. 0.2 M Sodium carbonate buffer, pH 9.5.

7. Sodium borohydride, 4 mg/mL (freshly prepared).

8. Sepharose CL-6B (Amersham Biosciences, Chalfont St. Giles, UK) or a similar gel filtration medium.

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

10. Bovine serum albumin (BSA).

0 0

Post a comment