C2 Immunostaining of Virally Infected CEF or QT6 Cells

1. PBS: PBS tablets are available from Sigma.

2. Triton X-100: available from Sigma.

3. Goat Serum: available from Gibco-BRL.

4. 3C2 hybridoma supernatant: available from the Developmental Studies Hybridoma Bank, Department of Biological Sciences, University of Iowa, Iowa City, IA.

5. Peroxidase-conjugated antimouse IgG, IgM secondary antibody: available from Jackson Laboratories, West Grove, PA.

6. 30% v/v Hydrogen peroxide: available from Sigma. Hydrogen peroxide can cause burns and is harmful by inhalation. Therefore adequate safety precautions should be taken.

7. 3.5% w/v Paraformaldehyde: dissolve 3.5 g paraformaldehyde in 100 mL of PBS (pH 7.4) by gently heating in a fume hood. Aliquot and store at -20°C. Paraformal-dehyde is extremely toxic. Therefore adequate safety precautions should be taken.

8. 0.5 mg/mL 3,3'-diaminobenzidine tetraaminobiphenyl (DAB) in PBS. DAB tablets (10 mg) are available from Sigma. Activate by adding 1 |L/mL of 30% (v/v) H2O2 to DAB just prior to use. DAB is extremely toxic. Therefore adequate safety precautions should be taken.

9. Nickel chloride: available from Sigma. Nickel chloride is a known mutagen and is extremely toxic. Therefore adequate safety precautions should be taken.

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