Protein ID Solutions Sample Prep Guide
Optional Depletion of High-Abundance Proteins with the Agilent Multiple Affinity Removal Column
Protocol for 4.6 x 50 mm column
1 If the LC system has been run with organic solvent, purge lines with water
for 15 to 20 min and inject water to remove organic solvent from the
autosampler injection loop. Do this step without a column.
2 Set up Buffer A and Buffer B as the only mobile phases.
3 Purge lines with Buffer A and Buffer B at a flow rate of 1.0 mL/min for 10
minutes without a column.
Be sure to purge all flow paths, including the autosampler injection loop,
to be absolutely sure there is no remaining organic solvent in the system.
Failure to do so will ruin the column.
4 Set up LC method (see
for details) and run two method blanks by
injecting 100 µL of Buffer A without a column.
N O T E
Consult column certificate of analysis to verify your column capacity.
For the mouse/rat column, the capacity range was determined with pooled Swiss Webster
mouse serum. Mouse serum protein concentrations can vary for other strains and your
injected sample size should be adjusted accordingly.
C A U T I O N
Do not expose columns to organic solvents (like alcohols, acetonitrile, etc.), strong
oxidizers, acids, or reducing agents, and other protein denaturing agents. If you do so,
the column will be ruined!
C A U T I O N
Ensure proper sample loop size in autosamplers!