Bio-Rad Aurum Serum Protein Mini Kit - Manual (Page 7)

Section 6
Protocol
Please read Section 5, “Guidelines for Aurum Serum Protein Mini Kit” before
proceeding.
1. Place an Aurum serum protein column in a 12 x 75 mm test tube and
allow the resin to settle for at least 5 min.
2. Remove the cap and break off the tip from the bottom of the Aurum
serum protein column. Return serum protein column to test tube.
3. Start gravity flow in the column and allow residual buffer to drain from the
column (approximately 2 min).
4. Once the residual buffer has drained, wash the column with 2 x 1 ml of
Aurum serum protein binding buffer using gravity flow. Allow each wash to
pass fully through the column and drain.
5. After the last wash, place the column in an empty 2.0 ml collection tube
and centrifuge for 20 sec at 10,000 x g in a microcentrifuge to dry resin
bed and frit. Do not overdry resin bed and frit. Discard the collection
tube.
6. Place a yellow column tip on the bottom of the column to stop any
residual buffer flow from column. Place the column in a clean 2.0 ml
collection tube labeled
“unbound”.
7. In a separate 2.0 ml collection tube, prepare sample to be purified by
diluting 60 µl of plasma or serum with 180 µl of Aurum serum protein
binding buffer. Mix by inverting tube several times.
8. Add 200 µl of the diluted serum sample to the top of the resin bed. Allow
sample to penetrate column matrix.
9. Gently vortex the column. Repeat at 5 and 10 min. Allow column to sit an
additional 5 min for a total incubation time of 15 min.
10. Remove yellow tip from column and return to 2.0 ml collection tube.
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