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Protein ID Solutions Sample Prep Guide
3
Optional Depletion of High-Abundance Proteins with Agilent Multiple Affinity Removal Spin Cartridges
Protocol for spin cartridges
1 Dilute and filter sample.
a Dilute serum to 200 µL. Specific instructions follow. Consult cartridge
certificate for actual sample capacity.
·
For human serum and High Capacity Multiple Affinity Removal Spin
Cartridge Hu-6HC, p/n 5188-5341:
Dilute 14 to 16 µL.
Example: If the recommended cartridge loading capacity on the
certificate is 14 µL of serum, dilute 14 µL of serum with 186 µL Buffer A
to a final volume of 200 µL.
·
For human serum and Multiple Affinity Removal Spin Cartridge Hu-6,
p/n 5188-5230:
Dilute 7 to 10 µL.
Example: If the recommended cartridge loading capacity on the
certificate is 8 µL of serum, dilute 8 µL of serum with 192 µL Buffer A to
a final volume of 200 µL.
·
For mouse/rat serum and Multiple Affinity Removal Spin Cartridge Ms-3,
p/n 5188-5289:
Dilute 25 to 30 µL.
Example: If the recommended cartridge loading capacity on the
certificate is 30 µL of serum, dilute 30 µL of serum with 170 µL Buffer A
to a final volume of 200 µL.
N O T E
Consult spin cartridge certificate of analysis to verify your cartridge capacity.
For the mouse/rat cartridge, the capacity range was determined with pooled Swiss
Webster mouse serum. Mouse serum protein concentrations can vary for other strains and
you should adjust your sample size accordingly.
C A U T I O N
Do not expose cartridges to organic solvents (like alcohols, acetonitrile, etc.), strong
oxidizers, acids, or reducing agents, and other protein denaturing agents. If you do so,
the cartridge will be ruined!
During use, never let the spin cartridge frits or resin bed run dry. If this happens, see
"Tips"
on page 35 for a cartridge re-wetting procedure.