Colony Screening

1. Nitrocellulose membranes (Schleicher and Schuell, pore size 0.45 Mm).

2. Peroxidase-labeled anti-mouse Ig antibodies (DAKO, Carpintiera, CA).

3. Incubation buffer (IB): TBST supplemented with 0.1% BSA, 1% fetal calf serum, and 1% horse serum.

4. Blocking solution: 3% low fat dried milk (from any supermarket) dissolved in TBST

5. Substrate buffer: mix 25.7 mL of 0.2 MNa2HPO4, 24.3 mL of 0.1 M citric acid and 50 mL of water (final pH 5.0).

6. DAB (diaminobenzidine dihydrochloride; Sigma, St Louis, MO) stock solution (80X: 32 mg/mL in water). This should be handled with care because it is a possible carcinogen.

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