Protein Expression and Purification Series - Bio-Rad
Protein Expression and Purification Series - Bio-Rad
Protein Expression and Purification Series - Bio-Rad
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<strong>Protein</strong> <strong>Expression</strong> <strong>and</strong> <strong>Purification</strong> <strong>Series</strong><br />
SDS-PAGE Electrophoresis Analysis of Purified Fractions<br />
Student Workstations<br />
Each student team requires the following items to analyze their purification samples via SDS-PAGE:<br />
Material Needed for Each Workstation Quantity<br />
Soluble PAGE sample 100 µl<br />
Flowthrough PAGE sample 100 µl<br />
Wash PAGE sample 100 µl<br />
Fraction A PAGE sample 100 µl<br />
Desalted A PAGE sample 50 µl<br />
Fraction B PAGE sample 100 µl<br />
Desalted B PAGE sample 50 µl<br />
Fraction C PAGE sample 100 µl<br />
Desalted C PAGE sample 50 µl<br />
Precision Plus <strong>Protein</strong> Dual Color st<strong>and</strong>ards 15 µl<br />
4–20% Mini-PROTEAN TGX precast gel 1<br />
Vertical electrophoresis chamber (per 1–4 gels) 1<br />
1x Tris/Glycine/SDS (TGS) running buffer (per electrophoresis chamber) 700–1,200 ml<br />
Power supply (200 V constant) to be shared between workstations 1<br />
Buffer dam (only required if running 1 gel/box) 1<br />
2–20 µl adjustable-volume micropipet <strong>and</strong> tips 1<br />
Staining tray 1<br />
<strong>Bio</strong>-Safe Coomassie stain (per gel) 50 ml<br />
Marking pen 1<br />
Common Workstation Quantity<br />
Water bath or heat block set to 95°C 1<br />
Microcentrifuge with variable speed setting >16,000 x g 1<br />
Water for gel washing <strong>and</strong> destaining (tap water is fine) 1 L<br />
Chapter 8: <strong>Purification</strong> Protocol for <strong>Bio</strong>Logic DuoFlow System<br />
193<br />
CHAPTER 8<br />
BIOLOGIC DUOFLOW<br />
PROTOCOL
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