The Green Fluorescent Protein Chromatography Kit can be used following the pGLO™ Bacterial Transformation Kit, which is purchased separately. A colony of transformed bacteria (produced using the pGLO Bacterial Transformation Kit) is placed in liquid cultures overnight, then lysed to release the bacteria's cellular contents. GFP is purified from the bacterial lysate using hydrophobic interaction chromatography (HIC) columns provided in this kit.
Features and Benefits
- Aligns with AP Biology Lab 6
- Fluorescent GFP allows real-time monitoring of the extraction and purification processes
- Uses bacteria transformed with the pGLO Bacterial Transformation Kit
How It Works
Green fluorescent protein is extremely hydrophobic compared to bacterial proteins. Unique characteristics of GFP enable it to be purified from bacterial cell proteins using HIC columns. When placed in a buffer containing a high concentration of salt, the HIC matrix selectively binds hydrophobic GFP molecules while allowing the bacterial proteins to pass through the column. Then, simply lowering the salt concentration of the buffer causes GFP to elute from the column in pure form. pGLO bacterial transformation and GFP purification together explore key processes used in biotechnology to produce and purify "designer proteins" with commercial or research value.
- Three-session laboratory activity, 45 min per session
- Provides sufficient materials for eight student workstations, up to four students per workstation