Bio-Rad Aurum Serum Protein Mini Kit - Manual (Page 9)

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Fig. 2. Aurum serum protein mini kit protocol overview
Aurum ™ Serum
Protein Mini Kit
Protocol Overview
Column Setup
1. Place serum protein column in a test tube
for 5 min to allow resin to settle.
2. Remove cap and break tip from column
and return to test tube to start gravity
flow in column.
3. Wash column with 1 ml of serum protein
binding buffer using gravity flow. Repeat.
4. Place column in empty 2.0 ml collection tube
and centrifuge for 20 sec at 10,000 x g to
dry resin bed. Discard collection tube.
5. Place a yellow column tip on bottom of column
and place into a clean 2.0 ml collection tube
labeled “unbound”.
Sample Binding and Purification
6. In a separate tube, prepare sample by diluting
60 µl serum or plasma with 180 µl of
serum protein binding buffer.
7. Add 200 µl of diluted serum to top of
resin bed in column.
8. Gently vortex column and repeat after
5 and 10 min. Allow column to sit
an additional 5 min.
Collection of Purified Samples
9. Remove yellow tip from column and return
to tube. Centrifuge column for 20 sec at
10,000 x g, collecting protein fraction in
“unbound” collection tube.
10. Using same collection tube, wash column
with 200 µl of serum protein binding buffer.
11. Centrifuge column for 20 sec at 10,000 x g,
collecting protein fraction in same “unbound”
collection tube. Discard serum protein column.
12. The combined fractions contain the albumin-
and IgG-depleted serum or plasma sample.
The sample is now ready for gel analysis.
1 ml
binding
buffer
60 µl
serum
200 µl
diluted
serum
200 µl
binding
buffer
180 µl
binding
buffer
Vortex
2x
3x
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