Bio-Rad BioLogic DuoFlow Pathfinder 80 System DuoFlow Chromatography System Starter Kit User Manual

Page 27

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3.9 Status Bar

At the bottom of the Manual screen is a status bar that is continually updated
with system parameters.

Section 4. Anion Exchange Separation of Protein Standards

The starter kit enables you to learn to use the DuoFlow Maximizer or Pathfinder
systems by programming and running 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.

4.1 Overview of the Procedure

Run Conditions

Buffer A1

50 mM Tris-HCl

Buffer A2

50 mM Tris base

Solution B1

deionized water

Solution B2

2.0 M NaCI

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 = 214nm), and 0.4 AUFS (λ = 304 nm)

Conductivity 100

mS/cm

pH

pH 7.1 to pH 9.1

General Procedure

Step 1

Prepare solutions A

1

, A

2

, B

1

, and B

2

Step 2

Prepare sample

Step 3

Install the UNO Q1 column

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