pGLO SDS-PAGE Extension

Image
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Overview

Extends the utility of the pGLO kit by using the green fluorescent protein expressed in bacteria to illustrate the central dogma of biology: DNA transcription, the expression of a protein, and the visualization of a trait.

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Description

Description

The pGLO Kit SDS-PAGE Extension is designed to be performed following the pGLO Bacterial Transformation Kit. The extension is designed to identify and visualize green fluorescent protein (GFP) from the proteome of transformed E. coli, which also contains thousands of other proteins. Because only the cloned GFP glows, students can use this phenotypic trait to quickly identify the GFP within the complex mix of proteins.

Features and Benefits

With this kit, students are able to:

  • Perform real biotechnology workflows
  • Identify proteins by conformation and size
  • Link gene induction to protein expression to protein identification
  • Understand chromophores and the basis of protein fluorescence
  • Extend the pGLO Bacterial Transformation Kit

How It Works

In the first part of the exercise, students use the pGLO Bacterial Transformation Kit to transfer pGLO plasmid encoding GFP into E. coli, a common prokaryotic organism used for DNA propagation and protein expression. Colonies of E. coli are qualitatively examined for fluorescence to determine whether the pGLO gene is being expressed. In the second part of the laboratory activity, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is employed to separate the entire repertoire of proteins expressed in E. coli, including the foreign GFP responsible for transferring the fluorescence trait.

More Information

  • Three-session laboratory activity, 45 min per session
  • Provides sufficient materials for eight student workstations, up to four students per workstation.

Specifications

Specifications

Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)
 
Laemmli sample buffer, 30 ml
1
Precision Plus Protein™ Kaleidoscope™ prestained standards, 50 µl
2
Dithiothreitol (DTT), 1 g
1
10x Tris-glycine-SDS electrophoresis buffer, 1 L
1
Bio-Safe™ Coomassie stain for proteins, 100 ml
1
Curriculum, including Teacher's Guide and Student Manual
1
Kit contains sufficient materials for 8 student workstations (2–4 students per workstation)
 
Laemmli sample buffer, 30 ml
1
Precision Plus Protein™ Kaleidoscope™ prestained standards, 50 µl
2
Dithiothreitol (DTT), 1 g
1
10x Tris-glycine-SDS electrophoresis buffer, 1 L
1
Bio-Safe™ Coomassie stain for proteins, 100 ml
1
Curriculum, including Teacher's Guide and Student Manual
1

Ordering

Ordering

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1660013EDU
Protein analysis classroom study kit, includes DTT, protein standards, Bio-Safe Coomassie stain, 10x TGS, Laemmli sample buffer, curriculum, for 32 students; education use only

Refills

Refills

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1660023EDU
Instruction manual for use with pGLO Kit SDS-PAGE Extension (#166-0013); education use only
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1610375EDU
500 µl, mixture of 10 multicolor recombinant proteins (10–250 kD), including three bands (25, 50, 75 kD) that match the reference bands of other standards, 50 applications; education use only
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1610732EDU
1 L, 10x premixed protein electrophoresis buffer, contains 25 mM Tris, 192 mM glycine, 0.1% SDS, pH 8.3; education use only

Documents

Documents

Number Description Options
6673
Map of AP-Aligned Bio-Rad Kits with Investigative Laboratories
M1660023
Instructor Guide, Protein Electrophoresis of GFP: A pGLO Bacterial Transformation Kit Extension