Bio-Rad BioLogic DuoFlow Pathfinder 80 System DuoFlow Chromatography System Starter Kit - Manual (Page 11)

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Section 2. Anion Exchange Separation of Protein Standards
The starter kit enables you to learn to use the DuoFlow system by programming
and running a separation of a premixed anion exchange standard containing
equine myoglobin, conalbumin, chicken ovalbumin and soybean trypsin inhibitor
using a 1.3 ml UNO Q1 column (catalog #720-0001). Equine myoglobin is not
retained on the UNO Q1 column and elutes in the void volume. Conalbumin,
chicken ovalbumin, and soybean trypsin inhibitor bind to the column and require
increased salt concentrations for elution. Separation requires approximately
6 minutes.
2.1 Overview of the Procedure
Run Conditions
• Buffer A 25 mM Tris-HCl, pH 8.1
• Buffer B 25 mM Tris-HCl, pH 8.1, 0.5 M NaCl
• Flow rate 4.00 ml/min
• Sample volume 50 µl
• UV detection 0.1 AUFS
• QuadTec detection 0.1 AUFS (
λ = 280 nm), 0.1 AUFS (λ = 260 nm),
1.0 AUFS ( λ = 214 nm), and 0.4 AUFS (λ = 304 nm)
• Conductivity 100 mS/cm
General Procedure
Step 1 Prepare buffer
Step 2 Prepare sample
Step 3 Install the UNO Q1 column
Step 4 Prime the workstation pumps and equilibrate the column
Step 5 Write a method
a. Program the instrument Setup
b. Program the method Protocol
c. Load sample into 50 µl loop
d. Select Run
e. Select Start
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