Protein ID Solutions Sample Prep Guide
19
Optional Depletion of High-Abundance Proteins with the Agilent Multiple Affinity Removal Column
2
Troubleshooting
N O T E
When not in use, store the LC with Buffer A in the lines. Buffer A contains sodium azide,
which inhibits bacterial growth. Periodically flush the LC system well with water to remove
salts which could damage pistons and seals.
Problem
Cause
Solution
High backpressure
Clogged inlet filters
Replace plugged frits (p/n 5185-5995) on both ends of column.
Remove particulates from samples with a spin filter prior to loading.
No bound fraction
peak
Insufficient exposure of
bound proteins to
Buffer B.
Check LC timetable to ensure enough column time exposure to Buffer B.
Abnormal peak
height
*
* Peak height of the bound fraction should be greater than that of the flow-through fraction.
Column may not have
been regenerated well
enough from previous
runs, resulting in lost
capacity.
Elute bound proteins with Buffer B for 3 additional minutes and
re-equilibrate the column with Buffer A.
Biological growth in
Buffer A
Check for signs of biological growth in the Buffer A reservoir.
Replace with fresh Buffer A for optimized column performance.
Summary :
Protein ID Solutions Sample Prep Guide 19 Optional Depletion of High-Abundance Proteins with the Agilent Multiple Affinity Removal Column 2 Troubleshooting N O T E When not in use, store the LC with Buffer A in the lines.
Tags :
buffer,column,bound,peak,fraction,proteins,growth,well,biological,enough,exposure,replace,check