You have performed an AAT assay on fraction ASIIdi and observe 0.00780 International Units of activity in the cuvet. The ASIIdi was diluted 200-fold and 100 L of this dilution was used in the assay. After completing the A₂₈₀measurements as described in the coursepack, the A₂₈₀of the final measurement, after 10 additions, was 0.112.

1. What is the A₂₈₀of fraction ASIIdi?

2. A fraction from the wash (fractions 1 - 20) step of the CM sephadex column has an A₂₈₀of 0.0660. How many fold would you need to dilute this fraction for the AAT assay (enter "1.00" if the fraction is assayed undiluted)?

3. Another fraction from the wash step of the CM sephadex column has an A₂₈₀of 0.0480. How many fold would you need to dilute this fraction to obtain an acceptable chart tracing (enter "1.00" if the fraction is assayed undiluted)?

4. A fraction from the elution step (fractions 21 - 40) of the CM sephadex column has an A₂₈₀of 0.460. How many fold would you need to dilute this fraction to obtain an acceptable chart tracing?

5. A fraction from the CM sephadex column has an A₂₈₀of 1.14. How many fold would you need to dilute this fraction to obtain an acceptable chart tracing?