Protein Expression and Purification Series - Bio-Rad

Protein Expression and Purification Series
Chapter 7: Purification Protocol for BioLogic LP System
Affinity Purification of GST-DHFR-His Using a BioLogic LP
System
Student Workstations
Each student team requires the following items to purify their GST-DHFR-His samples using affinity
chromatography:
Material Needed for Each Workstation Quantity
Soluble lysate fraction 5–7.5 ml
Either a BioScale Mini Profinity Ni-IMAC Cartridge 1
or or
An Econo-Column Glass column handpacked with 1 ml of Profinity IMAC Ni-Charged resin 1
10 ml syringe 1
Distilled water 500 ml
Buffer A 70 ml
Buffer B 40 ml
Collection tubes for fraction collector 32
Marking pen 1
Common Workstation Quantity
BioLogic LP system instrumentation, with sample loop installed 1–4
Preparing the Biologic LP for Use
Starting up the BioLogic LP system
Before a purification method can be run, you must take proper steps to evacuate all air from the system
and fill up the instrument lines with water to purge any storage buffer that might be in the tubing.
1. Make sure that there are no chromatography
columns connected to the system. The
column inlet tubing should be connected to the
column outlet tubing (Figure 7.1).
A B C
Figure 7.1. System connections without a column in place.
(A) Male luer fitting with barb (B) Female luer fitting with barb (C)
Flow path of the BioLogic LP system.
Chapter 7: Purification Protocol for BioLogic LP System
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CHAPTER 7
BIOLOGIC LP SYSTEM
PROTOCOL