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Star Republic: Guide for Biologists

Iodination protocol

Edited by Chang Zhu
1. Volatile 125I2 may be produced. Work under a chemical hood. Proper protection is also needed when working with 125I.

2. Dilute 2 mg of protein to 20 ml in 50 mM sodium phosphate (pH 7.4), 0.15 M NaCl.

3. Add 50 ml 0.5 M sodium phosphate (pH 7.0).

4. Add 1 mCi of Na125I.

5. Add 5 ml chloramine T (0.5 mg/ml in 50 mM sodium phosphate (pH 7.4), 0.15 M NaCl). Prepare chloramine solution immediately before use.

6. Incubate for 15 seconds to 5 minutes.

7. Stop the reaction by adding 10 ml Na2SO3 or Na2S3O5 (5 mg/ml).

8. Separate by gel filtration.