Using a pipette, add 2 mL of suspended DEAE SEPHAROSE (anion) resin to a chromatographic column. Repeat the same step to another column. Make sure the column valve is closed. Let it settle by gravity until you see a defined and settled top. 2. Open the column valve and let the preservative liquid slowly drip through until there remains only tiny bit on top of the column. Close the column valve. Repeat the same step to another column. 3. Add 10 mL of equilibration buffer and let it pass through the column (to wash out preservative). Discard as waste. Repeat the same step to another column. Close the valve when 8 mL of the equilibration buffer pass through the column. 4. Label 2 sets of test tubes 1-6. Label one set for lysozyme and the other set for albumin. 5. Using a micropipette, gently add 200 µL of lysozyme solution to the top of the first column (without disturbing the top of the bed). Open the column valve and allow the sample to "load" into the column. Collect the flow through into tube #1 for the first protein set. Close the column valve before the column runs dry. Repeat the same procedure on the second column by adding 200µL of albumin solution. 6. Add another 2 mL of equilibration buffer and let it pass through the column slowly. Collect the flow through into test tube # 2. 7. Repeat step 6, three more times for the column, collecting fractions slowly into test tubes #3, 4, 5 respectively. 8. Add 2 mL of elution (high salt) buffer to the top of the column. Open the valve. 9. As the elution slowly comes through, collect the final 2 mL sample in test tube #6. Close the column valve just before the column runs dry. 10. Test each fraction in your "run" with Bradford reagent to determine the presence of protein, by following the directions below. Blue color indicates the presence of protein. a) Turn on the spectrophotometer. Let it warm up 15 minutes. b) Add 3.0 mL of Bradford reagent to each of the test tubes 1-6 for each of the two protein sets. c) Cover each of the tubes with a tiny square of Parafilm. d) Invert each tube 3x to mix. Be gentle. Don't allow the solution to foam up. Don't mix up the Parafilm covers. Wait for 2 minutes before taking absorbance readings. e) Transfer the solutions to proper spec cuvettes (plastic) f) Make up a blank with 2.0 mL of equilibration buffer and 3.0 mL of Bradford reagent. Mix. g) Set the Spec to 595 nm. Collect the baseline using the blank following the instructions for the spectrophotometer. h) Wipe the outside of each spec tube and read the absorbance of each set of samples 1-6