Procedure

1 Fix the slide in equal parts methanol and acetone. Store at 4°C until stained.

2 Treat the slide with trypsin solution for 30 min at 37°C to remove interfering immunoglobulins. Paraffin sections do not need this treatment.

3 Transfer the slide to three changes of PBS, 1min each.

4 Incubate with monoclonal antibody in a moist chamber for 30min at RT.

5 Transfer to three changes of PBS for 1min each.

6 Incubate with diluted class specific monoclonal anti-mouse FITC conjugate for 30min at RT in a moist chamber.

7 Rinse in three changes of PBS for 1min each.

8 Mount under a coverslip with buffered glycerine.

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