Protein Expression and Purification Series - Bio-Rad

Protein Expression and Purification Series
Step 6: Check that the purified GST-DHFR-His has enzymatic activity.
Results: The “No Substrate Control Reaction” should show a fairly flat line with an absorbance around
0.2–0.4 (Fgure 8). The enzyme reaction should show a decrease in absorbance at 340 nm over the
150 seconds of reading time (Figure 9). The calculated reaction rate will vary depending on the amount of
GST-DHFR-His purified.
Absorbance at 340 nm
Figure 8. No substrate control enzymatic activity reaction.
Absorbance at 340 nm
Figure 9. GST-DHFR-His enzyme activity reaction.
Appendix B: Purification Results Analysis
No Substrate Control Reaction
Time in seconds
GST-DHFR-His Enzyme Reaction
Time in seconds
217
APPENDIX B
RESULTS ANALYSIS