Bio-Rad Bio-Scale™ MT High-Resolution Empty Columns - Manual (Page 10)

6 Bio-Scale MT Column Instruction Manual
5. Attach the column to a pump and start flow at 50% of the
desired operating flow rate.
6. When all the air is expelled from the column, increase the flow
rate to 20% higher than the desired operating flow rate for at
least 5 column volumes.
7. Stop the flow and lower the upper bed support via the lock nut
to remove any head space that might have formed between
the frit and the top of the bed.
8. Equilibrate the column at the desired flow rate prior to sample
application.
Slurry Packing
Slurry Preparation
Dry media should be swollen according to the manufacturer’s
instructions.
Always degas the buffers used for column packing. Adjust the
packing slurry to a 50% (V/V) volume using packing buffer.
Column Packing
1. Assemble the column according to the diagram in Figure 2
and seal the bottom outlet with Parafilm® laboratory film.
2. Fill about 25% of the column with buffer and add the packing
slurry to the desired level.
3. Uncap the bottom outlet and allow the slurry to settle. Remove
the film from the outlet, leaving some liquid above the settled
slurry.
4. Add more slurry to the desired level.
5. Without inserting the upper frit and distribution screen,
assemble the column and connect to a pump. Begin the
flow at 50% of the desired operating flow rate for 3 column
volumes.
6. Stop the pump, remove the upper bed support, and repeat the
above steps until a stable bed at the desired level is obtained.
7. Add 5 mm of buffer to the top of the bed and insert the upper
frit and distribution screen. Assemble the column.