Proliferation Assay Protocol

Tritium 3H Proliferation Assay Protocol

Tritium is a radioactive compound with a half life of 12.4 years. Its principal emission is 18.6 keV at maximum. It has a maximum ragne in air of 6 mm. No shield is required and monitoring for emission is through swipe tests counted through a liquid scintillator.

1. Incubate cells (e.g. cultured cells) with antibody for 1 day.

2. Count cells using a hemacytometer.

3. Transfer cells into a 96-well plate.

4. Each 0.1 mL of o.1 mCi/mL 3H thymidine to the appropriate well.

5. Label plates with radioactive tape

6. Incubate all cells at 37 celsius for 1 day

7. Harvest cells on glass-fiber disks using a cell harverster (Cells are lysed as they pass through filters. DNA will then stick to these filters, while the rest of the lysate will pass through. The amount of reactivity is then counted on each filter using a scintillation counter.)