Green Fluorescent Protein Chromatography Kit

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Overview

Lab Preparation Checklist
Lab Preparation Checklist
Kit contains sufficient materials for 8 student workstations. (2–4 students per workstation)
Ampicillin, lyophilized 1
Arabinose, lyophilized 1
Lysozyme, lyophilized 1
LB nutrient broth tablet 1
Binding buffer 1
Column equilibration buffer 1
Column wash buffer 1
Elution buffer 1
HIC chromatography columns 8
Inoculation loops, sterile 20
Disposable plastic transfer pipets 40
Micro test tubes, 2.0 ml, clear 30
Cell culture tubes, 15 ml, sterile 25
Sample collection tubes, 5 ml 25
Foam micro test tube holders 8
Curriculum including Teacher's Guide, Student Manual, and graphic Quick Guide 1
Required Accessories Not Included in Kit:
Transformed bacterial colonies from pGLO bacterial transformation kit  
UV lamps 4–8
Centrifuges 1–2
Recommended (Optional) Accessories:
Incubation oven  
Rocking platform  
Microwave oven  
PowerPoint Presentation for Educational Use
PowerPoint Presentation for Workshops and for Teaching

pGLO Transformation and Green Fluorescent Protein Chromatography Kits

This PowerPoint presentation is intended for educational use only. The presentation provides educators with background on the pGLO Transformation and Green Fluorescent Protein Chromatography Kits and includes instructions for using the kits.

pGLO Transformation and Green Fluorescent Protein Chromatography Kits Download PowerPoint presentation		 pGLO Transformation and Green Fluorescent Protein Chromatography Kits
GFP Chromatography Kit Workflow
GFP Chromatography Kit Workflow
Key Kit Features
Key Kit Features
  • Transform bacteria with a gene from a bioluminescent jellyfish (AP Lab 6)
  • Induce transformed bacteria to overexpress Green Fluorescent Protein
  • Purify GFP using chromatography
  • Complete in three 45 minute lab sessions
Curriculum Fit
Curriculum Fit
GFP Chromatography
GFP Chromatography

Chromatography 101. Chromatography to purify proteins of interest depends on a protein's chemical or physical properties, such as molecular weight, electrical charge, or solubility. Green Fluorescent Protein (GFP) is extremely hydrophobic compared to bacterial proteins. This unique characteristic of GFP enables the purification of GFP from bacterial cell proteins using hydrophobic interaction chromatography (HIC). 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 right 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 plate and a Series of HIC columns showing the separation of GFP.

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.

More Information

  • Three-session laboratory activity, 45 min per session
  • Provides sufficient materials for eight student workstations, up to four students per workstation
Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)  
Ampicillin, lyophilized 1
Arabinose, lyophilized 1
Lysozyme, lyophilized 1
LB nutrient broth tablet 1
Binding buffer 1
Column equilibration buffer 1
Column wash buffer 1
Elution buffer 1
HIC chromatography columns 8
Inoculation loops, sterile 20
Disposable plastic transfer pipets 40
Micro test tubes, 2.0 ml, clear 30
Cell culture tubes, 15 ml, sterile 25
Sample collection tubes, 5 ml 25
Foam micro test tube holders 8
Curriculum, including Teacher's Guide, Student Manual, and graphic Quick Guide 1
Green Fluorescent Protein Chromatography Kit

166-0005EDU
Protein purification classroom kit, includes lyophilized reagents, nutrient broth tablet, buffers, HIC columns, pipets, test tubes, curriculum, and more, for 32 students; education use only

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Literature
Number Description Options
3053 Green Fluorescent Protein Chromatography Kit, Biotechnology Explorer, Quick Guide Click to download
4006099 Instruction Manual, Green Fluorescent Protein (GFP) Purification Kit (pGLO Transformation Module 2), Biotechnology Explorer, Rev C Please login to download this document [ Add to Cart ]