28
Protein ID Solutions Sample Prep Guide
3
Optional Depletion of High-Abundance Proteins with Agilent Multiple Affinity Removal Spin Cartridges
Procedure summary
Figure 5
Procedure for multiple affinity removal of high-abundance proteins
5. Wash and collect flow-
through F2
8. Re-equilibration
3. Apply sample
4. Wash and collect flow-
through F1
6. Prepare for elution
7. Elute bound fraction
9. Analyze F1 and F2
200 µL diluted serum
Centrifuge 1.5 min at 100 × g
1. Dilute and filter sample
2. Prepare spin cartridge
4 mL Buffer A
400 µL Buffer A
Centrifuge 2.5 min at 100 × g
400 µL Buffer A
Centrifuge 2.5 min at 100 × g
2 mL Buffer B
Save or discard
4 mL Buffer A
Individually or combined
Concentrate first
Dilute. (See table on next page.)
Filter.
F1
F1
F2
F1 + F2
Summary :
Prepare spin cartridge 4 mL Buffer A 400 µL Buffer A Centrifuge 2.5 min at 100 × g 400 µL Buffer A Centrifuge 2.5 min at 100 × g 2 mL Buffer B Save or discard 4 mL Buffer A Individually or combined Concentrate first Dilute.
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