When bacteria are used to make medicinally useful proteins by transformation, the protein of interest must be separated from all of the other cellular proteins. In this experiment, the unique fluorescent properties of GFP and BFP are used as an assay during their purification from an E. coli extract. The column fractions containing GFP or BFP are identified by fluorescence and then purified. As an optional activity, purified protein fractions can be separated by SDS polyacrylamide gel electrophoresis (SDS-PAGE) to estimate the purity and size of the GFP and BFP proteins.
- For 6 lab groups
- Packing and running column 45 minutes; optional electrophoresis 60 minutes; staining 30 minutes; destaining 2 hours
Also required: Water bath, longwave UV lamp, ring stand and clamps, automatic micropipette, vertical gel electrophoresis apparatus, power supply, polyacrylamide gels (12%).
Delivery information: Kit includes instructions, columns and matrix, GFP and BFP extracts, buffer, protein gel reagents for optional activity.