Home Biochemistry Using the GELFREE 8100 Fractionation System for Molecular Weight-Based Fractionation with Liquid Phase Recovery
Biochemistry JoVE (Open Access) Citable · DOI

Using the GELFREE 8100 Fractionation System for Molecular Weight-Based Fractionation with Liquid Phase Recovery

DOI: 10.3791/1842-v
What you'll learn
  • Operate the GELFREE 8100 Fractionation System for molecular weight-based protein separation
  • Prepare complex protein samples and load cartridges correctly
  • Collect liquid-phase protein fractions and analyze results by PAGE
Protocol

The accompanying video describes the use of the GELFREE 8100 Fractionation System, which partitions complex protein samples on the basis of molecular weight and recovers the fractions in liquid phase. The video describes how the technology works, how it is used, and provides resultant data, with polyacrylamide gel electrophoresis analysis of fractionated bovine liver homogenate.

Difficulty
intermediate
Total time
~3–4 hours per sample (including run time and fraction collection)
Biosafety
BSL-1

Steps

1
Understand GELFREE 8100 technology and workflow

Learn the principles of molecular weight-based protein fractionation using gel electrophoresis in a tube format with liquid-phase recovery, and review the complete experimental workflow.

▶ 00:11
2
Prepare protein samples for fractionation

Process and prepare complex protein samples (e.g., bovine liver homogenate) to appropriate concentration and format for loading into the GELFREE cartridge.

▶ 00:48
3
Load sample into GELFREE 8100 cartridge

Insert the prepared protein sample into the cartridge following the system's specifications to ensure proper fractionation during the run.

▶ 01:53
4
Run cartridge and collect liquid fractions

Execute the GELFREE 8100 separation program and systematically collect molecular weight-fractionated protein samples in liquid phase as they elute from the system.

▶ 03:00
5
Analyze fractions using polyacrylamide gel electrophoresis

Perform PAGE analysis on collected fractions to visualize protein composition by molecular weight and confirm successful fractionation.

▶ 06:37
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